Polymorphs of sepiapterin and salts thereof

ABSTRACT

Disclosed are crystalline forms of sepiapterin free base selected from polymorphs A, B, C, D, E, F, and G, and combinations thereof, as well as crystalline polymorphs of salts of sepiapterin. Also disclosed are pharmaceutical compositions containing one or more such polymorphs and methods for preparing such polymorphs. Sepiapterin is useful in the treatment of a number diseases associated with low cellular levels of BH4, for example, phenylketonuria.

BACKGROUND OF THE INVENTION

Sepiapterin is a naturally occurring precursor of tetrahydrobiopterin(BH4), a naturally occurring essential cofactor of criticalintracellular enzymes to include but not limited to phenylalaninehydroxylase (PAH) (Kaufman, 1958), tyrosine hydroxylase (TH) (Nagatsu etal, 1964), tryptophan hydroxylase (TPH) (Ichiyama et al, 1970), nitricoxide synthases (NOS) (Kwon et al, 1989), (Mayer et al, 1991) andalkylglycerol monooxygenase (AGMO) (Tietz et al, 1964). Rapid conversionof sepiapterin to BH4 favoring accumulation of BH4 occurs via a two-stepreduction in the salvage pathway for BH4 synthesis (Sawabe, 2008). Asynthetic form of BH4 (e.g., sapropterin dihydrochloride) is used as atherapy for diseases associated with high plasma phenylalanine, such asphenylketonuria (PKU). PKU is an inborn error of metabolism causedpredominantly by mutations in the PAH gene. BH4 was also tested as atherapy for various central nervous symptoms associated with PKU andother diseases, but demonstrated limited effect, presumably due to theinability of BH4 to effectively cross the blood brain barrier (Klaimanet al, 2013; Grant et al, 2015).

Recent work has suggested that, compared with BH4, peripherallyadministered sepiapterin possesses greater permeability throughmembranes and as a result, can more readily access liver, kidney, andbrain cells. It is reported that sepiapterin is rapidly converted intoBH4 intracellularly via the tetrahydrobiopterin-salvage pathway, therebyelevating liver, kidney, and brain BH4 levels (Sawabe, 2008). As aresult, sepiapterin may serve as a useful therapeutic for diseasesassociated with low intracellular BH4 levels or with dysfunction ofvarious BH4 dependent metabolic pathways.

Sepiapterin herein is the S-enantiomer and has the formula (I):

It is known that sepiapterin has limited stability in solutions.Furthermore, certain forms of solid sepiapterin degrade under oxidativeconditions even at room temperature and in the presence of light.

Accordingly, there exists an unmet need for stable solid-state forms ofsepiapterin.

BRIEF SUMMARY OF THE INVENTION

The invention provides a solid form of sepiapterin free base, whereinthe solid form comprises an amorphous form of sepiapterin free base, asingle polymorph form of sepiapterin free base, a mixture of polymorphforms of sepiapterin free base, a salt of sepiapterin or a mixture ofsalts of sepiapterin, or a combination thereof, with the proviso thatthe solid form is not a pure polymorph of Form A or Form E sepiapterinfree base.

The invention also provides crystalline polymorph forms of salts ofsepiapterin, or a mixture thereof. As an example, the invention providesa crystalline polymorph form of sepiapterin hydrochloride salt.

It has now been surprisingly found that under certain conditions, newcrystalline forms of sepiapterin free base and acid salts are formed,which have advantageous utilities and properties. The invention thusprovides methods for preparing the various polymorphic forms.

The invention further provides pharmaceutical compositions comprisingone or more of these polymorphic forms.

In an aspect, the invention features a crystalline form sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 8.4°±0.5,16.9°±0.5, or 25.4°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry.In some embodiments, the crystalline form of sepiapterin has at leastone peak at diffraction angle 2θ (°) of 8.4°±0.5, 16.9°±0.5, and25.4°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry. In some embodiments, thecrystalline form of sepiapterin has at least one peak at diffractionangle 2θ (°) of 14.9°±0.5, or 34.1°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry. In some embodiments, the crystalline form of sepiapterinhas at least one peak at diffraction angle 2θ (°) of 8.4°±0.5,14.9°±0.5, 16.9°±0.5, 25.4°±0.5, and 34.1°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.

In some embodiments, the crystalline form of sepiapterin has the X-raypowder diffraction spectrum as shown in FIG. 1. In some embodiments, thecrystalline form of sepiapterin has an endothermic onset at about 195°C. in differential scanning calorimetry (DSC) profile.

In an aspect, the invention features crystalline form sepiapterin havingat least one peak at diffraction angle 2θ (°) of 5.7°±0.5, 7.8°±0.5, or25.4°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry. In some embodiments, thecrystalline form of sepiapterin has at least one peak at diffractionangle 2θ (°) of 5.7°±0.5, 7.8°±0.5, and 25.4°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry. In some embodiments, the crystalline form of sepiapterinhas at least one peak at diffraction angle 2θ (°) of 9.1°±0.5,11.5°±0.5, 15.3°±0.5, 16.0°±0.5, 20.1°±0.5, or 26.6°±0.5 as measured byX-ray diffractometry by irradiation with Cu Kα X-rays or calculated fromX-ray diffractometry. In some embodiments, the crystalline form ofsepiapterin has at least one peak at diffraction angle 2θ (°) of5.7°±0.5, 7.8°±0.5, 9.1°±0.5, 11.5°±0.5, 15.3°±0.5, 16.0°±0.5,20.1°±0.5, 25.4°±0.5, and 26.6°±0.5 as measured by X-ray diffractometryby irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.

In some embodiments, the crystalline form of sepiapterin has the X-raypowder diffraction spectrum as shown in FIG. 2. In some embodiments, thecrystalline form of sepiapterin has an endothermic onset at about 58°C., 102° C., 130° C., 156.5° C., or 168° C. in differential scanningcalorimetry (DSC) profile. In some embodiments, the crystalline form ofsepiapterin has an endothermic onset at about 58° C., 102° C., 130° C.,156.5° C., and 168° C. in differential scanning calorimetry (DSC)profile.

In an aspect, the invention features a crystalline form sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 8.9°±0.5,10.3°±0.5, or 26.0°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry.In some embodiments, the crystalline form of sepiapterin has at leastone peak at diffraction angle 2θ (°) of 8.9°±0.5, 10.3°±0.5, and26.0°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry. In some embodiments, thecrystalline form of sepiapterin has at least one peak at diffractionangle 2θ (°) of 10.9°±0.5, 17.8°±0.5, 24.9°±0.5, 26.7°±0.5, 26.8°±0.5,or 28.3°±0.5 as measured by X-ray diffractometry by irradiation with CuKα X-rays or calculated from X-ray diffractometry. In some embodiments,the crystalline form of sepiapterin has at least one peak at diffractionangle 20)(° of 8.9°±0.5, 10.3°±0.5, 10.9°±0.5, 17.8°±0.5, 24.9°±0.5,26.0°±0.5, 26.7°±0.5, 26.8°±0.5, and 28.3°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.

In some embodiments, the crystalline form of sepiapterin has the X-raypowder diffraction spectrum as shown in FIG. 3. In some embodiments, thecrystalline form of sepiapterin has an endothermic onset at about 43°C., 66° C., or 233° C. in differential scanning calorimetry (DSC)profile. In some embodiments, the crystalline form of sepiapterin has anendothermic onset at about 43° C., 66° C., and 233° C. in differentialscanning calorimetry (DSC) profile.

In an aspect, the invention features a crystalline form sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 9.7°±0.5,10.2°±0.5, or 11.3°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry.In some embodiments, the crystalline form of sepiapterin has at leastone peak at diffraction angle 2θ (°) of 9.7°±0.5, 10.2°±0.5, and11.3°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry. In some embodiments, thecrystalline form of sepiapterin has at least one peak at diffractionangle 2θ (°) of 14.0°±0.5, 14.6°±0.5, 19.9°±0.5, 22.2°±0.5, 25.3°±0.5,or 32.4°±0.5 as measured by X-ray diffractometry by irradiation with CuKα X-rays or calculated from X-ray diffractometry. In some embodiments,the crystalline form of sepiapterin has at least one peak at diffractionangle 20)(° of 9.7°±0.5, 10.2°±0.5, 11.3°±0.5, 14.0°±0.5, 14.6°±0.5,19.9°±0.5, 22.2°±0.5, 25.3°±0.5, and 32.4°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.

In some embodiments, the crystalline form of sepiapterin has the X-raypowder diffraction spectrum as shown in FIG. 4. In some embodiments, thecrystalline form of sepiapterin has an endothermic onset at about 113°C. or 196° C. in differential scanning calorimetry (DSC) profile. Insome embodiments, the crystalline form of sepiapterin has an endothermiconset at about 113° C. and 196° C. in differential scanning calorimetry(DSC) profile.

In an aspect, the invention features a composition including any of theforegoing crystalline forms of sepiapterin, or combinations thereof. Insome embodiments of the composition, the crystalline form of sepiapterinof any one of claims 1 to 27, is present in an amount of at least 90percent by weight of the composition.

In an aspect, the invention features a pharmaceutical compositionincluding any of the foregoing crystalline forms of sepiapterin. In someembodiments, the crystalline form of sepiapterin is formulated asparticles between 50 μm and 250 μm in size (e.g., less than 100 μm insize).

In an aspect, the invention features a method for preparing acrystalline form of sepiapterin including preparing a slurry of a firstcrystalline form of sepiapterin in water, acetone/water,isopropanol/isopropyl acetate, or tetrahydrofuran/n-hexane, isolatingthe solids from the slurry, and drying the solids. In some embodiments,the slurry of the first crystalline form of sepiapterin is stirred at25-75° C. for 6-72 hours. In some embodiments, the solids are dried at20-30° C. for 6-24 hours. In some embodiments, the solids are dried at40-60° C. for 5-10 hours. In some embodiments, the solids are dried atatmospheric pressure. In some embodiments, the solids are dried undervacuum.

In an aspect, the invention features a salt of sepiapterin. In someembodiments, the salt of sepiapterin is the methansulfonate salt, thenicotinate salt, the p-toluenesulfonate salt, the benzenesulfonate, thephosphate salt, the malonate salt, the tartrate salt, the gentisatesalt, the fumarate salt, the glycolate salt, the acetate salt, thesulfate salt, or the hydrochloride salt.

In an aspect, the invention features a crystalline form of a salt ofsepiapterin, wherein the crystalline form of a salt of sepiapterin is:

(a) a crystalline form of the methanesulfonate salt of sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 7.8°±0.5,23.5°±0.5, and/or 29.0°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry;

(b) a crystalline form of the methanesulfonate salt of sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 21.7°±0.5,26.0°±0.5, and/or 28.9°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry

(c) a crystalline form of the nicotinate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 9.5°±0.5, 9.9°±0.5, and/or24.5°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry;

(d) a crystalline form of the p-toluenesulfonate salt of sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 6.5°±0.5,15.1°±0.5, and/or 23.4°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry;

(e) a crystalline form of the benzenesulfonate salt of sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 6.5°±0.5,14.8°±0.5, and/or 19.6°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry;

(f) a crystalline form of the phosphate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 16.6°±0.5, 22.2°±0.5,and/or 25.6°±0.5 as measured by X-ray diffractometry by irradiation withCu Kα X-rays or calculated from X-ray diffractometry;

(g) a crystalline form of the malonate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 6.9°±0.5, 22.7°±0.5,and/or 23.8°±0.5 as measured by X-ray diffractometry by irradiation withCu Kα X-rays or calculated from X-ray diffractometry;

(h) a crystalline form of the tartrate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 7.3°±0.5, 14.2°±0.5,and/or 21.8°±0.5 as measured by X-ray diffractometry by irradiation withCu Kα X-rays or calculated from X-ray diffractometry;

(i) a crystalline form of the gentisate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 7.1°±0.5, 8.7°±0.5, and/or26.7°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry;

(j) a crystalline form of the fumarate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 11.3°±0.5, 24.0°±0.5,and/or 28.2°±0.5 as measured by X-ray diffractometry by irradiation withCu Kα X-rays or calculated from X-ray diffractometry;

(k) a crystalline form of the glycolate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 7.6°±0.5, 10.7°±0.5,and/or 24.0°±0.5 as measured by X-ray diffractometry by irradiation withCu Kα X-rays or calculated from X-ray diffractometry;

(l) a crystalline form of the acetate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 6.2°±0.5, 12.0°±0.5,and/or 18.1°±0.5 as measured by X-ray diffractometry by irradiation withCu Kα X-rays or calculated from X-ray diffractometry;

(m) a crystalline form of the sulfate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 5.1°±0.5, 7.8°±0.5, and/or23.0°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry; or

(n) a crystalline form of the sulfate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 7.8°±0.5, 8.8°±0.5, and/or24.1°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry.

In an aspect, the invention features a crystalline form of thehydrochloride salt of sepiapterin having at least one peak atdiffraction angle 2θ (°) of 7.8°±0.5, 12.9°±0.5, and/or 26.2°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry

In an aspect, the invention features a composition including any of theforegoing crystalline forms of a salt of sepiapterin. In someembodiments, the crystalline form of the salt of sepiapterin is presentin at least 90 percent by weight.

In an aspect, the invention features a pharmaceutical compositionincluding any of the foregoing crystalline forms of a salt ofsepiapterin and a pharmaceutically acceptable carrier. In someembodiments, the crystalline form of sepiapterin is formulated asparticles less than 100 μm in size.

In another aspect, the invention features a method for treating a BH4related disorder in a patient in need thereof, the method comprisingadministering to the patient an effective amount of any of the foregoingcrystalline forms of sepiapterin or pharmaceutical compositions. In someembodiments, the BH4-related disorder is a disease associated with lowintracellular BH4 levels or with dysfunction of various BH4 dependentmetabolic pathways including, but not limited to, primarytetrahydrobiopterin deficiency, GTPCH deficiency,6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, DHPR deficiency,sepiapterin reductase deficiency, dopamine responsive dystonia, SegawaSyndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12deficiency, Parkinson's Disease, depression due to Parkinson's Disease,impulsivity in Parkinson's patients, major depression, Autism spectrum,ADHD, schizophrenia, Bipolar disorder, cerebral ischemia, restless legsyndrome, Obsessive Compulsive Disorder, anxiety, aggression inAlzheimer's disease, cerebrovascular disorders, gastroparesis, spasmafter subarachnoidal hemorrhage, myocarditis, coronary vasospasm,cardiac hypertrophy, arteriosclerosis, hypertension, thrombosis,infections, endotoxin shock, hepatic cirrhosis, hypertrophic pyloricstenosis, gastric mucosal injury, pulmonary hypertension, renaldysfunction, impotence, and hypoglycemia. Thus, the various forms ofsepiapterin in accordance with the present invention can be administeredto a patient in an effective amount to obtain a treatment oramelioration of the disease or dysfunction.

In another aspect, the invention features a method of increasing BH4,serotonin, and/or dopamine levels (e.g., at least 5%, 10%, 20%, 30%,40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 1000% ormore) in a subject in need thereof, the method comprising administeringto the patient an effective amount of any of the foregoing crystallineforms of sepiapterin or pharmaceutical compositions.

In another aspect, the invention features a method of decreasingphenylalanine levels (e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 1000% or more) in a subjectin need thereof, the method comprising administering to the patient aneffective amount of any of the foregoing crystalline forms ofsepiapterin or a pharmaceutical compositions.

In another aspect, the invention features a method of increasing theactivity (e.g., at least 55%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,90%, 100%, 200%, 300%, 400%, 500%, 1000% or more) of phenylalaninehydroxylase, tyrosine hydroxylase, tryptophan hydroxylase, nitric oxidesynthase, and/or alkylglycerol monooxygenase in a subject, the methodcomprising administering to the patient an effective amount of any ofthe foregoing crystalline forms of sepiapterin or a pharmaceuticalcompositions.

In another aspect, the invention features a method of treatingphenylketonuria in a subject in need thereof, the method comprisingadministering to the patient an effective amount of any of the foregoingcrystalline forms of sepiapterin or pharmaceutical compositions.

Definitions

In this application, unless otherwise clear from context, (i) the term“a” may be understood to mean “at least one”; (ii) the term “or” may beunderstood to mean “and/or”; (iii) the terms “comprising” and“including” may be understood to encompass itemized components or stepswhether presented by themselves or together with one or more additionalcomponents or steps; and (iv) the terms “about” and “approximately” maybe understood to permit standard variation as would be understood bythose of ordinary skill in the art; and (v) where ranges are provided,endpoints are included.

As used herein, the term “administration” refers to the administrationof a composition (e.g., a compound or a preparation that includes acompound as described herein) to a subject or system.

Administration to an animal subject (e.g., to a human) may be by anyappropriate route. For example, in some embodiments, administration maybe bronchial (including by bronchial instillation), buccal, enteral,interdermal, intra-arterial, intradermal, intragastric, intramedullary,intramuscular, intranasal, intraperitoneal, intrathecal, intravenous,intraventricular, mucosal, nasal, oral, rectal, subcutaneous,sublingual, topical, tracheal (including by intratracheal instillation),transdermal, vaginal and vitreal.

As used herein, the term “BH4 related disorder,” refers to any diseaseor disorder that may derive a therapeutic benefit from modulation (e.g.,inhibition) of the level of BH4, e.g., phenylketonuria.

By “determining the level of a protein” is meant the detection of aprotein, or an mRNA encoding the protein, by methods known in the arteither directly or indirectly. “Directly determining” means performing aprocess (e.g., performing an assay or test on a sample or “analyzing asample” as that term is defined herein) to obtain the physical entity orvalue. “Indirectly determining” refers to receiving the physical entityor value from another party or source (e.g., a third party laboratorythat directly acquired the physical entity or value). Methods to measureprotein level generally include, but are not limited to, westernblotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA),radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surfaceplasmon resonance, chemiluminescence, fluorescent polarization,phosphorescence, immunohistochemical analysis, matrix-assisted laserdesorption/ionization time-of-flight (MALDI-TOF) mass spectrometry,liquid chromatography (LC)-mass spectrometry, microcytometry,microscopy, fluorescence activated cell sorting (FACS), and flowcytometry, as well as assays based on a property of a protein including,but not limited to, enzymatic activity or interaction with other proteinpartners. Methods to measure mRNA levels are known in the art.

An “effective amount” of a compound may vary according to factors suchas the disease state, age, sex, and weight of the individual, and theability of the compound to elicit the desired response. Atherapeutically effective amount encompasses an amount in which anytoxic or detrimental effects of the compound are outweighed by thetherapeutically beneficial effects. A therapeutically effective amountalso encompasses an amount sufficient to confer benefit, e.g., clinicalbenefit.

By “increasing the activity of phenylalanine hydroxylase,” is meantincreasing the level of an activity related to phenylalaninehydroxylase, or a related downstream effect. A non-limiting example ofincreasing an activity of phenylalanine hydroxylase is decreasing thelevel of phenylalanine. The activity level of phenylalanine hydroxylasemay be measured using any method known in the art.

By “level” is meant a level of a protein, or mRNA encoding the protein,as compared to a reference. The reference can be any useful reference,as defined herein. By a “decreased level” or an “increased level” of aprotein is meant a decrease or increase in protein level, as compared toa reference (e.g., a decrease or an increase by about 5%, about 10%,about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%,about 80%, about 85%, about 90%, about 95%, about 100%, about 150%,about 200%, about 300%, about 400%, about 500%, or more; a decrease oran increase of more than about 10%, about 15%, about 20%, about 50%,about 75%, about 100%, or about 200%, as compared to a reference; adecrease or an increase by less than about 0.01-fold, about 0.02-fold,about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, or less;or an increase by more than about 1.2-fold, about 1.4-fold, about1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold,about 20-fold, about 30-fold, about 40-fold, about 50-fold, about100-fold, about 1000-fold, or more). A level of a protein may beexpressed in mass/vol (e.g., g/dL, mg/mL, μg/mL, ng/mL) or percentagerelative to total protein or mRNA in a sample.

The term “pharmaceutical composition,” as used herein, represents acomposition containing a compound described herein formulated with apharmaceutically acceptable excipient, and manufactured or sold with theapproval of a governmental regulatory agency as part of a therapeuticregimen for the treatment of disease in a mammal. Pharmaceuticalcompositions can be formulated, for example, for oral administration inunit dosage form (e.g., a tablet, capsule, caplet, gelcap, suspension,solution, or syrup); for topical administration (e.g., as a cream, gel,lotion, or ointment); for intravenous administration (e.g., as a sterilesolution free of particulate emboli and in a solvent system suitable forintravenous use); or in any other pharmaceutically acceptableformulation.

A “pharmaceutically acceptable excipient,” as used herein, refers anyingredient other than the compounds described herein (for example, avehicle capable of suspending or dissolving the active compound) andhaving the properties of being substantially nontoxic andnon-inflammatory in a patient. Excipients may include, for example:antiadherents, antioxidants, binders, coatings, compression aids,disintegrants, dyes (colors), emollients, emulsifiers, fillers(diluents), film formers or coatings, flavors, fragrances, glidants(flow enhancers), lubricants, preservatives, printing inks, sorbents,suspensing or dispersing agents, sweeteners, and waters of hydration.Exemplary excipients include, but are not limited to: ascorbic acid,butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate(dibasic), calcium stearate, colloidal silicon dioxide, croscarmellose,croscarmellose sodium, crosslinked polyvinyl pyrrolidone, citric acid,crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropylcellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate,maltitol, mannitol, methionine, methylcellulose, methyl paraben,microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone,povidone, pregelatinized starch, propyl paraben, retinyl palmitate,shellac, silicon dioxide, sodium carboxymethyl cellulose, sodiumcitrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid,sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, andxylitol.

As used herein, the term “pharmaceutically acceptable salt” means anypharmaceutically acceptable salt of the compound of formula (I). Forexample, pharmaceutically acceptable salts of any of the compoundsdescribed herein include those that are within the scope of soundmedical judgment, suitable for use in contact with the tissues of humansand animals without undue toxicity, irritation, allergic response andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example,pharmaceutically acceptable salts are described in: Berge et al., J.Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts:Properties, Selection, and Use, (Eds. P. H. Stahl and C. G. Wermuth),Wiley-VCH, 2008. The salts can be prepared in situ during the finalisolation and purification of the compounds described herein orseparately by reacting a free base group with a suitable organic acid.

The compounds of the invention may have ionizable groups so as to becapable of preparation as pharmaceutically acceptable salts. These saltsmay be acid addition salts involving inorganic or organic acids or thesalts may, in the case of acidic forms of the compounds of the inventionbe prepared from inorganic or organic bases. Frequently, the compoundsare prepared or used as pharmaceutically acceptable salts prepared asaddition products of pharmaceutically acceptable acids or bases.Suitable pharmaceutically acceptable acids and bases and methods forpreparation of the appropriate salts are well-known in the art. Saltsmay be prepared from pharmaceutically acceptable non-toxic acids andbases including inorganic and organic acids and bases.

Representative acid addition salts include acetate, adipate, alginate,ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate,butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, toluenesulfonate, undecanoate, and valeratesalts. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, and magnesium, as well as nontoxicammonium, quaternary ammonium, and amine cations, including, but notlimited to ammonium, tetramethylammonium, tetraethylammonium,methylamine, dimethylamine, trimethylamine, triethylamine, andethylamine.

By a “reference” is meant any useful reference used to compare proteinor mRNA levels. The reference can be any sample, standard, standardcurve, or level that is used for comparison purposes. The reference canbe a normal reference sample or a reference standard or level. A“reference sample” can be, for example, a control, e.g., a predeterminednegative control value such as a “normal control” or a prior sampletaken from the same subject; a sample from a normal healthy subject,such as a normal cell or normal tissue; a sample (e.g., a cell ortissue) from a subject not having a disease; a sample from a subjectthat is diagnosed with a disease, but not yet treated with a compound ofthe invention; a sample from a subject that has been treated by acompound of the invention; or a sample of a purified protein (e.g., anydescribed herein) at a known normal concentration. By “referencestandard or level” is meant a value or number derived from a referencesample. A “normal control value” is a pre-determined value indicative ofnon-disease state, e.g., a value expected in a healthy control subject.Typically, a normal control value is expressed as a range (“between Xand Y”), a high threshold (“no higher than X”), or a low threshold (“nolower than X”). A subject having a measured value within the normalcontrol value for a particular biomarker is typically referred to as“within normal limits” for that biomarker. A normal reference standardor level can be a value or number derived from a normal subject nothaving a disease or disorder (e.g., cancer); a subject that has beentreated with a compound of the invention. In preferred embodiments, thereference sample, standard, or level is matched to the sample subjectsample by at least one of the following criteria: age, weight, sex,disease stage, and overall health. A standard curve of levels of apurified protein, e.g., any described herein, within the normalreference range can also be used as a reference.

As used herein, the term “subject” or “patient” refers to any organismto which a composition in accordance with the invention may beadministered, e.g., for experimental, diagnostic, prophylactic, and/ortherapeutic purposes. Typical subjects include any animal (e.g., mammalssuch as mice, rats, rabbits, non-human primates, and humans). A subjectmay seek or be in need of treatment, require treatment, be receivingtreatment, be receiving treatment in the future, or be a human or animalwho is under care by a trained professional for a particular disease orcondition.

As used herein, the terms “treat,” “treated,” or “treating” mean boththerapeutic treatment and prophylactic or preventative measures whereinthe object is to prevent or slow down (lessen) an undesiredphysiological condition, disorder, or disease, or obtain beneficial ordesired clinical results. Beneficial or desired clinical resultsinclude, but are not limited to, alleviation of symptoms; diminishmentof the extent of a condition, disorder, or disease; stabilized (i.e.,not worsening) state of condition, disorder, or disease; delay in onsetor slowing of condition, disorder, or disease progression; ameliorationof the condition, disorder, or disease state or remission (whetherpartial or total), whether detectable or undetectable; an ameliorationof at least one measurable physical parameter, not necessarilydiscernible by the patient; or enhancement or improvement of condition,disorder, or disease. Treatment includes eliciting a clinicallysignificant response without excessive levels of side effects. Treatmentalso includes prolonging survival as compared to expected survival ifnot receiving treatment.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Methods and materials aredescribed herein for use in the present disclosure; other, suitablemethods and materials known in the art can also be used. The materials,methods, and examples are illustrative only and not intended to belimiting. All publications, patent applications, patents, sequences,database entries, and other references mentioned herein are incorporatedby reference in their entirety. In case of conflict, the presentspecification, including definitions, will control.

The details of one or more embodiments of the invention are set forth inthe description below. Other features, objects, and advantages of theinvention will be apparent from the description and from the claims.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING(S)

FIG. 1 shows the X-ray diffraction diagram of the crystalline Form B ofsepiapterin free base.

FIG. 2 shows the X-ray diffraction diagram of the crystalline Form C ofsepiapterin free base.

FIG. 3 shows the X-ray diffraction diagram of the crystalline Form D ofsepiapterin free base.

FIG. 4 shows the X-ray diffraction diagram of the crystalline Form F ofsepiapterin free base.

FIG. 5 shows the X-ray diffraction diagram of the crystalline Form G ofsepiapterin free base.

FIG. 6 shows an overlay of the X-ray diffraction diagrams of thecrystalline Form 1 hydrochloride salt of sepiapterin and of the startingsepiapterin free base used in the preparation of the hydrochloride salt.

FIG. 7 shows an overlay of the X-ray diffraction diagrams of thecrystalline Form 1 methanesulfonate salt, of Form 2 methanesulfonatesalt, of Form 3 methanesulfonate salts of sepiapterin and of thestarting sepiapterin free base used in the preparation of themethanesulfonate salts.

FIG. 8 shows an overlay of the X-ray diffraction diagrams of thecrystalline nicotinate salt of sepiapterin, of nicotinic acid, and ofthe starting sepiapterin free base used in the preparation of thenicotinate salt.

FIG. 9 shows an overlay of the X-ray diffraction diagrams of thecrystalline p-toluenesulfonate salt of sepiapterin, of p-toluenesulfonic acid, and of the starting sepiapterin free base used in thepreparation of the p-toluenesulfonate salt.

FIG. 10 shows an overlay of the X-ray diffraction diagrams of thecrystalline benzenesulfonate salt of sepiapterin, of benzene sulfonicacid, and of the starting sepiapterin free base used in the preparationof the benzenesulfonate salt.

FIG. 11 shows an overlay of the X-ray diffraction diagrams of thecrystalline phosphate salt of sepiapterin and of the startingsepiapterin free base used in the preparation of the phosphate salt.

FIG. 12 shows an overlay of the X-ray diffraction diagrams of thecrystalline malonate salt of sepiapterin, of malonic acid, and of thestarting sepiapterin free base used in the preparation of the malonatesalt.

FIG. 13 shows an overlay of the X-ray diffraction diagrams of thecrystalline L-tartrate salt of sepiapterin, of L-tartaric acid, and ofthe starting sepiapterin free base used in the preparation of theL-tartrate salt.

FIG. 14 shows an overlay of the X-ray diffraction diagrams of thecrystalline gentisate salt of sepiapterin, of gentisic acid, and of thestarting sepiapterin free base used in the preparation of the gentisatesalt.

FIG. 15 shows an overlay of the X-ray diffraction diagrams of thecrystalline fumarate salt of sepiapterin, of fumaric acid, and of thestarting sepiapterin free base used in the preparation of the fumaratesalt.

FIG. 16 shows an overlay of the X-ray diffraction diagrams of thecrystalline glycolate salt of sepiapterin, of glycolic acid, and of thestarting sepiapterin free base used in the preparation of the glycolatesalt.

FIG. 17 shows an overlay of the X-ray diffraction diagrams of thecrystalline acetate salt of sepiapterin and of the starting sepiapterinfree base used in the preparation of the acetate salt.

FIG. 18 shows an overlay of the X-ray diffraction diagrams of thecrystalline sepiapterin Form 1 sulfate salt, of the crystallinesepiapterin Form 2 sulfate salt, and of the starting sepiapterin freebase used in the preparation of the sulfate salts.

FIG. 19 shows an overlay of the X-ray diffraction diagrams of thecrystalline forms of sepiapterin Form A before and after a grinding andsieving process and confirms the physical form stability thereof togrinding and sieving.

FIG. 20 shows an overlay of the X-ray diffraction diagrams of thecrystalline forms of sepiapterin Form F before and after a grinding andsieving process and confirms the stability thereof to grinding andsieving.

FIG. 21 shows an overlay of the X-ray diffraction diagrams of thecrystalline forms of sepiapterin Form D before and after a grinding andsieving process along with sepiapterin Form D and Form F as references.This Figure demonstrates the potential instability of Form D to grindingand sieving.

FIG. 22 shows the X-ray diffraction diagram of the crystalline Form E ofsepiapterin free base.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a solid form of sepiapterin, wherein thesolid form comprises an amorphous form, a crystalline polymorph form, amixture of amorphous and/or crystalline polymorph forms, a salt ofsepiapterin, or a combination thereof, with the proviso that the solidform of sepiapterin is not a pure polymorph Form A and/or pure polymorphForm E.

In an embodiment, in the solid form of sepiapterin the mixture comprisesat least one of crystalline polymorph Form B, C, D, F, and G ofsepiapterin.

In an embodiment, the solid form comprises at least one crystallinesepiapterin free base selected from polymorph Forms B, C, D, F, and Gand crystalline polymorph A or E or both crystalline polymorphs A and E.

The polymorphic forms of sepiapterin free base as well as thepolymorphic forms of the salts of sepiapterin may be characterized byany suitable method for studying solid state materials. In anembodiment, the polymorphic forms are characterized by X-ray powderDiffractometry (XRD). The XRD peak positions are expressed as the 20°.In the X-ray diagram, the angle of refraction 2θ is plotted on thehorizontal axis (x-axis) and the relative peak intensity(background-corrected peak intensity) on the vertical (y-axis). X-raypowder diffraction patterns are measured on, or using instrumentscomparable to, a PANalytical Empyrean X-ray powder diffractometer withCu Kα radiation source (Kα1 radiation, wavelength λ=1.54060 Angstrom,Kα2 radiation, wavelength 1.544426 Angstrom; Kα2/Kα1 intensity ratio:0.50). The optical density of the peaks on the film is proportional tothe light intensity. The film is scanned using a peak scanner.

As it relates to any of the peaks of X-ray powder diffraction set forththroughout this application, “about” refers to ±0.1, particularly ±0.05,and more particularly ±0.02 of the 20 values in degrees.

In an embodiment, the crystalline polymorph Form B of sepiapterin ischaracterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 8.4, about 16.9, and about 25.4°.

In a particular embodiment, the crystalline polymorph Form B ofsepiapterin is characterized by an X-ray powder diffraction patternobtained by irradiation with Cu Kα X-rays having peaks expressed as 20at least at about 8.4, about 14.9, about 16.9, about 25.4, and about34.1°.

In an embodiment, the crystalline polymorph Form C of sepiapterin ischaracterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 5.7, about 7.8, and about 25.4°.

In a particular embodiment, the crystalline sepiapterin polymorph Form Cis characterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 5.7, about 7.8, about 9.1, about 11.5, about 15.3, about 16.0,about 20.1, about 25.4, and about 26.6°.

In an embodiment, the crystalline sepiapterin polymorph Form D ischaracterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 8.9, about 10.3, and about 26.0°.

In a particular embodiment, the crystalline sepiapterin polymorph Form Dis characterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 8.9, about 10.3, about 10.9, about 17.8, about 24.9, about 26.0,about 26.7, about 26.8, and about 28.3°.

In an embodiment, the crystalline sepiapterin polymorph Form F ischaracterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 9.7, about 10.2, and about 11.3°.

In a particular embodiment, the crystalline sepiapterin polymorph Form Fis characterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 9.7, about 10.2, about 11.3, about 14.0, about 14.6, about 19.9,about 22.2, about 25.3, and about 32.4°.

In an embodiment, the crystalline sepiapterin polymorph Form G ischaracterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 10.0, about 10.6, and about 25.7°.

In a particular embodiment, the crystalline sepiapterin polymorph Form Gis characterized by an X-ray powder diffraction pattern obtained byirradiation with Cu Kα X-rays having peaks expressed as 20 at least atabout 10.0, about 10.6, about 11.2, about 15.3, about 15.9, about 22.8,about 24.4, about 25.0, about 25.7, and about 26.6°.

In an embodiment, the solid form comprises at least one crystalsepiapterin free base selected from polymorph forms B, C, D, F, and G;selected from polymorph forms B, C, and D; selected from polymorph formsB, C, and F; selected from polymorph forms D, F, and G; as well as anybinary, ternary, or quaternary combinations of the polymorph forms. Thesolid forms indicated above could further include polymorph A and/or E.

In an embodiment, polymorph Form B, C, D, or G, or a combinationthereof, is present in the solid form in an amount of at least 90percent by weight of the solid form.

In certain embodiments, the crystalline sepiapterin free base is presentin at least 70 percent or more by weight, at least 80 percent or more byweight, and preferably at least 90 percent or more by weight, based onthe weight of the sepiapterin free base.

The crystalline Form A of sepiapterin free base is characterized by anX-ray powder diffraction pattern obtained by irradiation with Cu KαX-rays having peaks expressed as 20 at least at about 4.7°, about 7.4°,about 9.5°, about 11.3°, about 15.6°, about 26.2°, and about 27.2°.

FIG. 19 shows the X-ray diffraction diagram of Form A of sepiapterinfree base. The most intense peak in the X-ray diffraction diagram isobserved at an angle of refraction 2θ of about 7.4°. The crystallineForm A is characterized by the 20 peak positions of at least about 4.7°,about 7.4°, about 9.5°, about 11.3°, about 15.6°, about 26.2°, and about27.2°. In an essentially pure material, crystal Form A of sepiapterinfree base, peaks can be observed at angles of refraction 2θ as set forthin Table 1.

TABLE 1 Position [2θ°] Relative Intensity 4.7 47.76 7.4 100.00 9.5 33.5411.3 19.31 12.4 8.49 13.4 3.60 14.2 8.24 15.6 15.08 16.4 11.97 17.6 8.3518.4 5.03 19.8 9.18 21.5 5.44 24.4 5.56 26.2 35.37 27.2 19.11 28.9 5.93

The crystalline Form B of sepiapterin free base is characterized bypeaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ of at least about 8.4, about 16.9, and about 25.4.

FIG. 1 shows the X-ray diffraction diagram of crystalline Form B ofsepiapterin free base. The most intense peak in the X-ray diffractiondiagram is observed at an angle of refraction 2θ of about 8.4°. Thecrystalline Form B is characterized by refractions at angles ofrefraction 2θ of at least about 8.4°, about 14.9°, about 16.9°, about25.4°, and about 34.1°. In an essentially pure material of the crystalForm B of sepiapterin free base, peaks can be observed at angles ofrefraction 2θ as set forth in Table 2.

TABLE 2 Position [2θ°] Relative Intensity 8.4 100.00 14.9 2.34 16.910.70 25.4 84.90 34.1 3.00

The crystalline Form C of sepiapterin free base is characterized bypeaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ of at least at about 5.7°, about 7.8°, and about 25.4°.

FIG. 2 shows the X-ray diffraction diagram of crystalline Form C ofsepiapterin free base. The most intense peak in the X-ray diffractiondiagram is observed at an angle of refraction 2θ of at least about 7.8°.Crystalline Form C is characterized by refractions at angles ofrefraction 2θ of at least about 5.7°, about 7.8°, about 9.1°, about11.5°, about 15.3°, about 16.0°, about 20.1°, about 25.4°, and about26.6°. In an essentially pure material of Form C of sepiapterin freebase, peaks can be observed at angles of refraction 2θ as set forth inTable 3.

TABLE 3 Position [2θ°] Relative Intensity 5.7 48.91 7.8 100.00 9.1 59.4910.4 8.72 11.5 24.53 12.9 8.50 14.8 9.24 15.3 12.53 16.0 14.09 17.2 7.2218.2 4.25 19.2 5.78 20.1 14.54 21.5 6.47 22.9 6.85 23.7 4.80 25.4 65.6826.6 14.53 27.4 8.39 31.5 3.74 34.2 4.36

The crystal Form D of sepiapterin free base is characterized by peaks inthe X-ray diffraction diagram observed at least at an angle ofrefraction 2θ of about 8.9°, about 10.3°, and about 26.0°.

FIG. 3 shows the X-ray diffraction diagram of crystal Form D ofsepiapterin free base. The most intense peak in the X-ray diffractiondiagram is observed at an angle of refraction 2θ of at least about 8.9°.The crystal Form D is characterized by refractions at angles ofrefraction 2θ of at least about 8.9°, about 10.3°, about 10.9°, about17.8°, about 24.9°, about 26.0°, about 26.7°, about 26.8°, and about28.3°. In an essentially pure material of Form D of sepiapterin freebase, peaks can be observed at angles of refraction 2θ as set forth inTable 4.

TABLE 4 Position [2θ°] Relative Intensity 8.9 100.00 10.3 49.92 10.919.96 11.6 2.15 13.6 2.99 14.2 3.45 14.8 2.35 15.4 2.59 16.4 1.55 17.22.33 17.8 6.24 19.6 2.62 20.1 2.28 20.5 3.09 20.8 2.27 21.3 3.60 22.34.79 23.7 4.31 24.9 5.19 26.0 41.94 26.7 8.58 26.8 9.17 27.4 3.98 28.34.75 28.7 6.60 29.8 3.03 31.8 2.72 33.0 2.03 35.5 1.57 37.1 1.09

The crystalline Form E of sepiapterin free base is characterized by anX-ray powder diffraction pattern obtained by irradiation with Cu KαX-rays having peaks expressed as 20 at least at about 6.0°, about 10.6°,about 12.1°, about 15.9°, about 20.8°, and about 24.6°.

FIG. 22 shows the X-ray diffraction diagram of Form E of sepiapterinfree base. The most intense peak in the X-ray diffraction diagram isobserved at an angle of refraction 2θ of at least about 6.0°. Thecrystalline Form E is characterized by refractions at angles ofrefraction 2θ of at least about 6.0°, about 10.6°, about 12.1°, about15.9°, about 20.9°, and about 24.6°. In an essentially pure form, forcrystalline Form E of sepiapterin free base, peaks can be observed atangles of refraction 2θ as set forth in Table 5.

TABLE 5 Position [2θ°] Relative Intensity 6.0 100.00 10.6 20.78 12.131.95 15.9 12.83 18.1 3.39 20.9 11.63 22.1 2.79 24.6 8.28 26.1 0.88 28.17.33 28.9 3.77 32.1 3.57 37.0 1.03

The crystalline Form F of sepiapterin free base is characterized bypeaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ of at least about 9.7°, about 10.2°, and about 11.3°.

FIG. 4 shows the X-ray diffraction diagram of the F form of sepiapterinfree base. The most intense peak in the X-ray diffraction diagram isobserved at an angle of refraction 2θ of at least about 10.2°. The Fform is characterized by refractions at angles of refraction 2θ of atleast about 9.7°, about 10.2°, about 11.3°, about 14.0°, about 14.6°,about 19.9°, about 22.2°, about 25.3°, and about 32.4°. In anessentially pure F form of sepiapterin free base, peaks can be observedat angles of refraction 2θ as set forth in Table 6.

TABLE 6 Position [2θ°] Relative Intensity 9.7 98.27 10.2 100.00 11.322.47 14.0 5.01 14.6 12.36 19.9 5.63 21.1 3.72 22.2 5.37 22.7 4.04 24.52.99 25.3 17.65 27.2 3.10 32.4 5.29 36.7 2.72

The crystalline form G of sepiapterin free base is characterized bypeaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ of at least about 10.0°, about 10.6°, and about 25.7°.FIG. 5 shows the X-ray diffraction diagram of the crystalline Form G ofsepiapterin free base obtained at 120° C. The most intense peak in theX-ray diffraction diagram is observed at an angle of refraction 2θ of atleast about 10.0°. More broadly, the G-crystal form is characterized byrefractions at angles of refraction 2θ of at least about 10.0°, about10.6°, about 11.2°, about 15.3°, about 15.9°, about 22.8°, about 24.4°,about 25.0°, about 25.7°, and about 26.6°. In an essentially purematerial of the G-crystal form of sepiapterin free base, peaks can beobserved at angles of refraction 2θ as set forth in Table 7.

TABLE 7 Position [2θ°] Relative Intensity 5.3 8.30 6.9 4.54 10.0 100.0010.6 69.64 11.2 6.59 13.5 7.52 15.3 26.59 15.9 26.43 16.0 23.41 16.94.28 18.6 13.02 19.3 11.90 20.1 7.22 20.8 11.01 22.8 16.77 23.5 19.6024.4 41.45 25.0 23.99 25.7 65.40 26.6 39.64 27.6 13.04 28.7 6.55 30.814.76 32.2 9.63 33.7 5.16 37.5 5.80

In the context of stating that crystalline Form A of sepiapterin freebase exhibits an X-ray diffraction diagram essentially as in FIG. 19,the term “essentially” means that at least the major peaks of thediagram depicted in FIG. 19, i.e., those having a relative peakintensity of more than 20%, especially more than 30%, as compared to themost intense peak in the diagram, have to be present.

Alternatively, or in addition, crystalline Form A of sepiapterin freebase is characterized by a DSC curve showing endothermal peaks at 82.8°C. and 179.8° C.

In the context of stating that crystalline Form B of sepiapterin freebase exhibits an X-ray diffraction diagram essentially as in FIG. 1, theterm “essentially” means that at least the major peaks of the diagramdepicted in FIG. 1, i.e. those having a relative peak intensity of morethan 20%, especially more than 30%, as compared to the most intense peakin the diagram, have to be present.

Alternatively, or in addition, the crystalline Form B of sepiapterinfree base is characterized by a DSC curve showing a melting event at195.2° C.

In a preferred embodiment, an essentially pure crystalline Form B ofsepiapterin free base shows the X-ray diffraction diagram indicated inFIG. 1.

In another preferred embodiment, crystalline Form B of sepiapterin freebase shows an X-ray diffraction diagram of the type shown in FIG. 1, inwhich the relative peak intensities of each peak do not deviate by morethan 10% from the relative peak intensities in the diagram shown in FIG.1, especially an X-ray diffraction diagram identical to that shown inFIG. 1.

In the context of stating that crystalline Form C of sepiapterin freebase exhibits an X-ray diffraction diagram essentially as in FIG. 2, theterm “essentially” means that at least the major peaks of the diagramdepicted in FIG. 2, i.e., those having a relative peak intensity of morethan 20%, especially more than 30%, as compared to the most intense peakin the diagram, have to be present.

Alternatively, the crystalline Form C of sepiapterin free base ischaracterized by a DSC curve showing five endothermal peaks at 58.3° C.,101.8° C., 129.8° C., 156.5° C., and 168.3° C.

In one preferred embodiment, the essentially pure crystalline Form C ofsepiapterin free base shows the X-ray diffraction diagram indicated inFIG. 2.

In another preferred embodiment, the crystalline Form C of sepiapterinfree base shows an X-ray diffraction diagram of the type shown in FIG.2, in which the relative peak intensities of each peak do not deviate bymore than 10% from the relative peak intensities in the diagram shown inFIG. 2, especially an X-ray diffraction diagram identical to that shownin FIG. 2.

In the context of stating that the crystalline Form D of sepiapterinfree base exhibits an X-ray diffraction diagram essentially as in FIG.3, the term “essentially” means that at least the major peaks of thediagram depicted in FIG. 3, i.e., those having a relative peak intensityof more than 20%, especially more than 30%, as compared to the mostintense peak in the diagram, have to be present.

Alternatively, the crystalline Form D of sepiapterin free base ischaracterized by a DSC curve showing three endotherms at 42.7° C., 66.3°C., and 232.9° C.

In one preferred embodiment, the essentially pure crystalline Form D ofsepiapterin free base shows the X-ray diffraction diagram indicated inFIG. 3.

In another preferred embodiment, the crystalline Form D of sepiapterinfree base shows an X-ray diffraction diagram of the type shown in FIG.3, in which the relative peak intensities of each peak do not deviate bymore than 10% from the relative peak intensities in the diagram shown inFIG. 3, especially an X-ray diffraction diagram identical to that shownin FIG. 3.

In the context of stating that the crystalline Form E of sepiapterinfree base exhibits an X-ray diffraction diagram essentially as in FIG.22, the term “essentially” means that at least the major peaks of thediagram depicted in FIG. 22, i.e. those having a relative peak intensityof more than 20%, especially more than 30%, as compared to the mostintense peak in the diagram, have to be present.

Alternatively, the crystalline Form E of sepiapterin free base ischaracterized by a DSC curve showing two endothermal peaks at 112.9° C.and 195.8° C.

In the context of stating that the crystalline Form F of sepiapterinfree base exhibits an X-ray diffraction diagram essentially as in FIG.4, the term “essentially” means that at least the major peaks of thediagram depicted in FIG. 4, i.e., those having a relative peak intensityof more than 20%, especially more than 30%, as compared to the mostintense peak in the diagram, have to be present.

Alternatively, the crystalline Form F of sepiapterin free base ischaracterized by a DSC curve showing two endotherms at 71.6° C. and233.4° C.

In one preferred embodiment, the essentially pure crystalline Form F ofsepiapterin free base shows the X-ray diffraction diagram indicated inFIG. 4.

In another preferred embodiment, the crystalline Form F of sepiapterinfree base shows an X-ray diffraction diagram of the type shown in FIG.4, in which the relative peak intensities of each peak do not deviate bymore than 10% from the relative peak intensities in the diagram shown inFIG. 4, especially an X-ray diffraction diagram identical to that shownin FIG. 4.

In the context of stating that the crystalline Form G of sepiapterinfree base exhibits an X-ray diffraction diagram essentially as in FIG.5, the term “essentially” means that at least the major peaks of thediagram depicted in FIG. 5, i.e., those having a relative peak intensityof more than 20%, especially more than 30%, as compared to the mostintense peak in the diagram, have to be present.

In one preferred embodiment, the essentially pure crystalline Form G ofsepiapterin free base shows the X-ray diffraction diagram indicated inFIG. 5.

In another preferred embodiment, the crystalline Form G of sepiapterinfree base shows an X-ray diffraction diagram of the type shown in FIG.5, in which the relative peak intensities of each peak do not deviate bymore than 10% from the relative peak intensities in the diagram shown inFIG. 5, especially an X-ray diffraction diagram identical to that shownin FIG. 5.

The invention also provides a crystalline form of sepiapterinhydrochloride salt.

In an embodiment, the crystalline hydrochloride salt is characterized byan X-ray powder diffraction pattern obtained by irradiation with Cu KαX-rays having peaks expressed as 20 at least at about 7.8°, about 12.9°,and about 26.2°.

The invention further provides a crystalline polymorph form of a salt ofsepiapterin. In certain embodiments, the invention provides acrystalline polymorph form of a salt of sepiapterin, wherein the salt isa salt of sepiapterin with sulfuric acid, p-toluene sulfonic acid,methane sulfonic acid, benzene sulfonic acid, malonic acid, tartaricacid (e.g., L-tartaric acid), phosphoric acid, gentisic acid, fumaricacid, glycolic acid, acetic acid, or nicotinic acid.

In particular embodiments, the crystalline polymorph salt is selectedfrom the group consisting of:

crystalline Form 1 methanesulfonate salt characterized by an X-raypowder diffraction pattern obtained by irradiation with Cu Kα X-rayshaving peaks expressed as 20 at least at about 7.8°, about 23.5°, andabout 29.0°;

crystalline Form 2 methanesulfonate salt characterized by an X-raypowder diffraction pattern obtained by irradiation with Cu Kα X-rayshaving peaks expressed as 20 at least at about 7.9°, about 23.4°, andabout 28.9°;

crystalline Form 3 methanesulfonate salt characterized by an X-raypowder diffraction pattern obtained by irradiation with Cu Kα X-rayshaving peaks expressed as 20 at least at about 21.7°, about 26.0°, andabout 28.9°;

crystalline nicotinate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 9.5°, about 9.9°, and about 24.5°;

crystalline p-toluenesulfonate salt characterized by an X-ray powderdiffraction pattern obtained by irradiation with Cu Kα X-rays havingpeaks expressed as 20 at least at about 6.5°, about 15.1°, and about23.4°;

crystalline benzenesulfonate salt characterized by an X-ray powderdiffraction pattern obtained by irradiation with Cu Kα X-rays havingpeaks expressed as 20 at least at about 6.5°, about 14.8°, and about19.6°;

crystalline phosphate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 16.6°, about 22.2°, and about 25.6°;

crystalline malonate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 6.9°, about 22.7°, and about 23.8°;

crystalline tartrate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 7.3°, about 14.2°, and about 21.8°;

crystalline gentisate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 7.1°, about 8.7°, and about 26.7°;

crystalline fumarate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at about 11.3°, about 24.0°, and about 28.2°;

crystalline glycolate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 7.6°, about 10.7°, and about 24.0°;

crystalline acetate salt characterized by an X-ray powder diffractionpattern obtained by irradiation with Cu Kα X-rays having peaks expressedas 20 at least at about 6.2°, about 12.0°, and about 18.1′;

crystalline Form 1 sulfate salt characterized by an X-ray powderdiffraction pattern obtained by irradiation with Cu Kα X-rays havingpeaks expressed as 20 at least at about 5.1°, about 7.8°, and about23.0°; and

crystalline Form 2 sulfate salt characterized by an X-ray powderdiffraction pattern obtained by irradiation with Cu Kα X-rays havingpeaks expressed as 20 at least at about 7.8°, about 8.8°, and about24.1°.

The crystalline hydrochloride salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ of at least at about 7.8°, about 12.9°, and about26.2°.

FIG. 6 shows the X-ray diffraction diagram of the crystallinehydrochloride salt of sepiapterin free base. The most intense peak inthe X-ray diffraction diagram is observed at an angle of refraction 2θof at least about 7.8°. In an essentially pure material of thecrystalline hydrochloride salt of sepiapterin free base, peaks can beobserved at angles of refraction 2θ as set forth in Table 8.

TABLE 8 Position [2θ°] Relative Intensity 7.8 100.00 8.9 6.89 12.9 58.5615.6 8.52 17.9 25.23 19.2 5.48 21.1 10.97 23.6 25.15 25.2 22.66 26.245.91 27.6 32.94 30.3 10.50 31.7 7.83 34.2 8.87 36.7 3.67

The crystalline Form 1 methanesulfonate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 7.8°, about 23.5°, and about29.0°.

FIG. 7 shows the X-ray diffraction diagram of the crystalline Form 1methanesulfonate salt of sepiapterin free base. The most intense peak inthe X-ray diffraction diagram is observed at an angle of refraction 2θof at least about 23.5°. In an essentially pure material of thecrystalline Form 1 methanesulfonate salt of sepiapterin free base, peakscan be observed at angles of refraction 2θ as set forth in Table 9.

TABLE 9 Position [2θ°] Relative Intensity 7.9 21.77 11.7 8.20 13.7 8.5215.7 4.79 16.6 5.34 18.0 5.66 19.8 2.10 20.3 5.36 20.9 2.43 22.3 4.2522.7 2.15 23.5 100.00 24.7 3.69 25.6 2.70 26.8 1.79 27.2 1.68 28.3 2.7529.0 57.60 29.8 5.18 30.5 1.37 32.2 4.66 33.0 1.64 36.5 1.29

The crystalline Form 2 methanesulfonate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 7.9°, about 23.4°, and about28.9°.

FIG. 7 shows the X-ray diffraction diagram of the crystalline Form 2methanesulfonate salt of sepiapterin free base. The most intense peak inthe X-ray diffraction diagram is observed at an angle of refraction 2θof at least about 23.5°. In an essentially pure material of thecrystalline Form 2 methanesulfonate salt of sepiapterin free base, peakscan be observed at angles of refraction 2θ as set forth in Table 10.

TABLE 10 Position [2θ°] Relative Intensity 7.9 100.00 11.0 21.32 12.122.02 13.5 79.87 15.7 11.87 17.8 9.81 19.7 10.93 21.3 26.79 23.4 96.1324.1 24.88 24.3 22.10 25.5 9.45 26.0 11.27 27.6 7.63 28.9 95.64 31.24.39 36.1 6.65

The crystalline Form 3 methanesulfonate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 21.7°, about 26.0°, and about28.9°.

FIG. 7 shows the X-ray diffraction diagram of the crystalline Form 3methanesulfonate salt of sepiapterin free base. The most intense peak inthe X-ray diffraction diagram is observed at an angle of refraction 2θof at least about 26.0°. In an essentially pure material of thecrystalline Form 3 methanesulfonate salt of sepiapterin free base, peakscan be observed at angles of refraction 2θ as set forth in Table 11.

TABLE 11 Position [2θ°] Relative Intensity 8.2 47.29 10.8 56.14 12.616.34 13.2 15.90 14.0 24.39 15.0 12.03 15.9 16.20 18.2 22.97 20.1 25.5320.5 14.97 21.3 22.70 21.7 71.48 22.2 11.40 23.6 46.37 24.8 44.00 25.59.08 26.1 100.00 27.3 3.52 28.9 68.42 31.2 4.49 32.1 6.48 34.8 5.95 35.61.67 39.1 2.91

The crystalline nicotinate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 9.5°, about 9.9°, and about24.5°.

FIG. 8 shows the X-ray diffraction diagram of the crystalline nicotinatesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 24.5°. In an essentially pure material of the crystallinenicotinate salt of sepiapterin free base, peaks can be observed atangles of refraction 2θ as set forth in Table 12.

TABLE 12 Position [2θ°] Relative Intensity 9.5 10.29 9.9 53.95 11.5 9.3112.0 11.76 14.7 14.20 15.9 17.61 17.5 7.53 19.0 5.37 20.8 5.88 21.3 6.1221.7 7.20 23.2 34.05 24.5 100.00 25.2 12.90 28.0 8.51 31.1 5.39 32.34.52 33.4 8.02 35.1 5.05

The crystalline p-toluenesulfonate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 6.5°, about 15.1°, and about23.4°.

FIG. 9 shows the X-ray diffraction diagram of the crystallinep-toluenesulfonate salt of sepiapterin free base. The most intense peakin the X-ray diffraction diagram is observed at an angle of refraction2θ of at least about 6.5°. In an essentially pure material of thep-toluenesulfonate salt of sepiapterin free base, peaks can be observedat angles of refraction 2θ as set forth in Table 13.

TABLE 13 Position [2θ°] Relative Intensity 6.5 100.00 12.9 1.79 14.31.39 15.1 15.36 16.2 5.33 18.4 8.96 19.6 3.06 20.2 4.86 21.8 2.23 22.52.95 23.1 7.99 23.4 9.14 24.5 1.81 26.0 2.48 27.0 4.49 27.3 3.93 28.15.31 28.4 5.59 28.8 2.05 30.6 2.24 31.0 1.98 32.6 1.82

The crystalline benzenesulfonate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 6.5°, about 14.8°, and about19.6°.

FIG. 10 shows the X-ray diffraction diagram of the crystallinebenzenesulfonate salt of sepiapterin free base. The most intense peak inthe X-ray diffraction diagram is observed at an angle of refraction 2θof at least about 6.5°. In an essentially pure material of thebenzenesulfonate salt of sepiapterin free base, peaks can be observed atangles of refraction 2θ as set forth in Table 14.

TABLE 14 Position [2θ°] Relative Intensity 4.9 5.90 6.5 100.00 14.816.73 17.8 4.23 19.6 7.98 21.5 2.49 23.7 3.46 24.5 3.84 26.1 3.29

The crystalline phosphate salt of sepiapterin free base is characterizedby peaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ at least at about 16.6°, about 22.2°, and about 25.6°.

FIG. 11 shows the X-ray diffraction diagram of the crystalline phosphatesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 25.6°. In an essentially pure material of the crystallinephosphate salt of sepiapterin free base, peaks can be observed at anglesof refraction 2θ as set forth in Table 15.

TABLE 15 Position [2θ°] Relative Intensity 5.5 4.41 8.1 1.21 8.9 2.2110.3 1.79 10.8 5.80 15.3 1.84 16.6 8.35 17.7 1.95 20.3 1.40 21.2 1.6122.2 9.77 23.1 1.74 25.6 100.00 30.8 6.31 31.1 4.85 33.5 0.73 36.0 1.70

The crystalline malonate salt of sepiapterin free base is characterizedby peaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ at least at about 6.9°, about 22.7°, and about 23.8°.

FIG. 12 shows the X-ray diffraction diagram of the crystalline malonatesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 6.9°. In an essentially pure material of the crystalline malonatesalt of sepiapterin free base, peaks can be observed at angles ofrefraction 2θ as set forth in Table 16.

TABLE 16 Position [2θ°] Relative Intensity 6.9 100.00 8.4 13.11 10.67.62 16.4 5.63 17.8 9.73 19.3 8.96 20.1 9.99 22.2 10.50 22.7 20.52 23.834.02 24.5 5.82 25.5 24.50 26.6 4.00 27.3 6.96 29.8 5.38 33.1 12.08

The crystalline L-tartrate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 7.3°, about 14.2°, and about21.8°.

FIG. 13 shows the X-ray diffraction diagram of the crystallineL-tartrate salt of sepiapterin free base. The most intense peak in theX-ray diffraction diagram is observed at an angle of refraction 2θ of atleast about 6.9°. In an essentially pure material of the crystallineL-tartrate salt of sepiapterin free base, peaks can be observed atangles of refraction 2θ as set forth in Table 17.

TABLE 17 Position [2θ°] Relative Intensity 7.4 100.00 10.1 47.99 14.282.76 14.7 27.06 19.1 21.16 20.2 29.91 21.8 85.30 22.1 53.68 23.9 85.3024.9 19.26 25.5 28.45 26.8 18.58 29.7 21.59 31.6 10.10 32.9 22.18

The crystalline gentisate salt of sepiapterin free base is characterizedby peaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ at least at about 7.1°, about 8.7°, and about 26.7°.

FIG. 14 shows the X-ray diffraction diagram of the crystalline gentisatesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 7.1°. In an essentially pure material of the crystalline gentisatesalt of sepiapterin free base, peaks can be observed at angles ofrefraction 2θ as set forth in Table 18.

TABLE 18 Position [2θ°] Relative Intensity 5.7 17.29 7.1 100.00 8.742.69 10.4 3.94 11.3 11.69 12.1 4.13 14.3 21.10 16.0 6.46 16.4 5.94 17.05.85 17.6 7.93 19.1 8.27 20.20 3.47 20.7 2.90 21.5 3.37 23.6 2.69 24.44.50 26.7 52.20 27.1 35.49 28.2 8.74 28.9 4.31 29.9 2.62 31.4 2.99 34.41.28 35.8 3.54 37.6 0.57

The crystalline fumarate salt of sepiapterin free base is characterizedby peaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ at least at about 11.3°, about 24.0°, and about 28.2°.

FIG. 15 shows the X-ray diffraction diagram of the crystalline fumaratesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 24.0°. In an essentially pure material of the crystalline fumaratesalt of sepiapterin free base, peaks can be observed at angles ofrefraction 2θ as set forth in Table 19.

TABLE 19 Position [2θ°] Relative Intensity 6.1 6.43 7.7 5.40 11.4 53.6211.9 33.37 14.2 8.03 16.5 6.70 18.3 13.86 19.0 6.68 20.7 10.02 21.3 7.0222.8 24.68 24.0 100.00 28.3 33.26 32.7 6.35 36.0 3.28 38.5 6.02

The crystalline glycolate salt of sepiapterin free base is characterizedby peaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ at least at about 7.6°, about 10.7°, and about 24.0°.

FIG. 16 shows the X-ray diffraction diagram of the crystalline glycolatesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 7.6°. In an essentially pure material of the crystalline glycolatesalt of sepiapterin free base, peaks can be observed at angles ofrefraction 2θ as set forth in Table 20.

TABLE 20 Position [2θ°] Relative Intensity 4.8 6.23 7.6 100.00 10.368.06 10.7 70.69 15.3 36.51 18.2 24.25 18.7 27.26 19.9 2.66 21.2 17.1124.0 96.62 24.4 18.44 28.8 47.57 30.3 7.43 32.5 4.42 33.3 7.49 34.3 5.2136.3 7.37

The crystalline acetate salt of sepiapterin free base is characterizedby peaks in the X-ray diffraction diagram observed at an angle ofrefraction 2θ at least at about 6.2°, about 12.0°, and about 18.1°.

FIG. 17 shows the X-ray diffraction diagram of the crystalline acetatesalt of sepiapterin free base. The most intense peak in the X-raydiffraction diagram is observed at an angle of refraction 2θ of at leastabout 6.2°. In an essentially pure material of the crystalline acetatesalt of sepiapterin free base, peaks can be observed at angles ofrefraction 2θ as set forth in Table 21.

TABLE 21 Position [2θ°] Relative Intensity 6.2 100.00 10.2 23.29 12.071.59 18.1 31.27 21.1 20.29 24.2 14.92 25.2 23.03 27.3 13.30 29.1 12.95

The crystalline Form 1 sulfate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 5.1°, about 7.8°, and about23.0°.

FIG. 18 shows the X-ray diffraction diagram of the crystalline Form 1sulfate salt of sepiapterin free base. The most intense peak in theX-ray diffraction diagram is observed at an angle of refraction 2θ of atleast about 5.1°. In an essentially pure material of the crystallineForm 1 sulfate salt of sepiapterin free base, peaks can be observed atangles of refraction 2θ as set forth in Table 22.

TABLE 22 Position [2θ°] Relative Intensity 5.1 100.00 6.8 3.33 7.8 43.4810.2 15.92 15.7 18.13 17.2 8.33 18.7 6.49 19.8 5.19 21.3 5.52 23.0 19.0523.5 8.29 24.2 5.59 24.8 17.44 25.7 4.97 26.7 10.38 28.7 11.49 30.4 2.8831.0 3.67

The crystalline Form 2 sulfate salt of sepiapterin free base ischaracterized by peaks in the X-ray diffraction diagram observed at anangle of refraction 2θ at least at about 7.8°, about 8.8°, and about24.1°.

FIG. 18 shows the X-ray diffraction diagram of the crystalline Form 2sulfate salt of sepiapterin free base. The most intense peak in theX-ray diffraction diagram is observed at an angle of refraction 2θ of atleast about 8.8°. In an essentially pure material of the crystallineForm 2 sulfate salt of sepiapterin free base, peaks can be observed atangles of refraction 2θ as set forth in Table 23.

TABLE 23 Position [2θ°] Relative Intensity 5.0 4.71 7.9 72.24 8.8 100.0014.5 19.26 15.7 59.40 16.1 8.69 17.2 14.82 17.7 10.89 19.3 9.92 20.29.60 23.7 15.38 24.2 43.88 25.0 11.44 26.8 16.81 28.7 16.07 29.4 13.8431.3 17.14 31.7 7.26 35.7 5.75

In the context of stating that the crystalline hydrochloride salt ofsepiapterin free base exhibits an X-ray diffraction diagram essentiallyas in FIG. 6, the term “essentially” means that at least the major peaksof the diagram depicted in FIG. 6, i.e. those having a relative peakintensity of more than 20%, especially more than 30%, as compared to themost intense peak in the diagram, have to be present. Alternatively, thecrystalline hydrochloride salt of sepiapterin free base is characterizedby a DSC curve showing an endotherm at 225.9° C.

In one preferred embodiment, the essentially pure crystallinehydrochloride salt of sepiapterin free base shows the X-ray diffractiondiagram indicated in FIG. 6.

In another preferred embodiment, the crystalline hydrochloride salt ofsepiapterin free base shows an X-ray diffraction diagram of the typeshown in FIG. 6, in which the relative peak intensities of each peak donot deviate by more than 10% from the relative peak intensities in thediagram shown in FIG. 6, especially an X-ray diffraction diagramidentical to that shown in FIG. 6.

In the context of stating that the crystalline salt forms of sepiapterinfree base, such as the crystalline form 1 methanesulfonate salt,crystalline form 2 methanesulfonate salt, crystalline form 3methanesulfonate salt, crystalline nicotinate salt, crystallinep-toluenesulfonate salt, crystalline benzenesulfonate salt, crystallinephosphate salt, crystalline malonate salt, crystalline L-tartrate salt,crystalline gentisate salt, crystalline fumarate salt, crystallineglycolate salt, crystalline acetate salt, crystalline form 1 sulfatesalt, and crystalline form 2 sulfate salt, exhibits an X-ray diffractiondiagram such as essentially as in FIGS. 7-18, respectively, the term“essentially” means that at least the major peaks of the diagramdepicted in FIGS. 7-18, i.e., those having a relative peak intensity ofmore than 20%, especially more than 30%, as compared to the most intensepeak in the diagram, have to be present.

In preferred embodiments, the essentially pure crystalline hydrochloridesalt of sepiapterin free base shows the X-ray diffraction diagramindicated in FIG. 6.

In another preferred embodiment, the crystalline form 1 methanesulfonatesalt, crystalline form 2 methanesulfonate salt, crystalline form 3methanesulfonate salt, crystalline nicotinate salt, crystallinep-toluenesulfonate salt, crystalline benzenesulfonate salt, crystallinephosphate salt, crystalline malonate salt, crystalline L-tartrate salt,crystalline gentisate salt, crystalline fumarate salt, crystallineglycolate salt, crystalline acetate salt, crystalline form 1 sulfatesalt, and crystalline form 2 sulfate salt of sepiapterin free base showsX-ray diffraction diagrams of the type shown in FIGS. 7-18, in which therelative peak intensities of each peak do not deviate by more than 10%from the relative peak intensities in the diagram shown in FIGS. 7-18,especially an X-ray diffraction diagram identical to that shown in FIGS.7-18, respectively.

Alternatively, the crystalline form 1 methanesulfonate salt ofsepiapterin free base is characterized by a DSC curve showing twoendotherms at 186.0° C. and 229.1° C.;

the crystalline form 2 methanesulfonate salt of sepiapterin free base ischaracterized by a DSC curve showing three endotherms at 75.5° C.,182.6° C., and 234.9° C.;

the crystalline form 3 methanesulfonate salt of sepiapterin free base ischaracterized by a DSC curve showing two endotherms at 195.1° C. and240.1° C.;

the crystalline nicotinate salt of sepiapterin free base ischaracterized by a DSC curve showing an endotherm at 221.9° C.;

the crystalline p-toluenesulfonate salt of sepiapterin free base ischaracterized by a DSC curve showing three endotherms at 77.2° C.,202.4° C. and 260.2° C.;

the crystalline benzenesulfonate salt of sepiapterin free base ischaracterized by a DSC curve showing two endotherms at 202.3° C. and265.5° C.;

the crystalline phosphate salt of sepiapterin free base is characterizedby a DSC curve showing three endotherms at 125.9° C., 152.1° C., and157.6° C.;

the crystalline malonate salt of sepiapterin free base is characterizedby a DSC curve showing a melting event at 115.8° C.;

the crystalline L-tartrate salt of sepiapterin free base ischaracterized by a DSC curve showing two endotherms at 97.2° C. and160.6° C.;

the crystalline gentisate salt of sepiapterin free base is characterizedby a DSC curve showing three endotherms at 70.5° C., 128.2° C., and184.7° C.;

the crystalline fumarate salt of sepiapterin free base is characterizedby a DSC curve showing two endotherms at 114.3° C. and 229.7° C.;

the crystalline glycolate salt of sepiapterin free base is characterizedby a DSC curve showing two endotherms at 133.9° C. and 147.7° C.;

the crystalline acetate salt of sepiapterin free base is characterizedby a DSC curve showing two endotherms at 146.1° C. and 175.4° C.; and

the crystalline form 1 sulfate salt of sepiapterin free base ischaracterized by a DSC curve showing three endotherms at 94.5° C.,158.3° C., and 209.9° C.

In any of the above embodiments, the crystalline sepiapterin free baseor a crystalline polymorph form of a salt of sepiapterin can occur as ananhydrate (e.g., without having any bound water or solvent or hydrationor solvation) or as a hydrate, a partial hydrate (e.g., hemihydrate,sesquihydrate, and the like), as a dihydrate, a trihydrate, or the like,wherein the crystalline form binds a water of hydration or a solventmolecule associated with the crystalline form of sepiapterin or saltthereof. In an embodiment, crystalline sepiapterin Form B occurs as ananhydrate. In an embodiment, crystalline sepiapterin Form C occurs as amonohydrate or as a sesquihydrate. In an embodiment, crystallinesepiapterin Form D occurs as a monohydrate or as a sesquihydrate. In anembodiment, crystalline sepiapterin Form F occurs as a monohydrate or asa hemihydrate. In an embodiment, crystalline sepiapterin Form G occursas an anhydrate.

In an embodiment, the invention provides a method for preparingcrystalline Form D of sepiapterin. The method comprises preparing aslurry of sepiapterin in a liquid, wherein the liquid is water,acetone/water, isopropanol/isopropyl acetate, ortetrahydrofuran/n-hexane, and isolating sepiapterin Form D from theslurry. Preferably, the liquid is water. The sepiapterin Form D can beisolated using any suitable isolation method, for example, bycentrifugation or by filtration. Preferably, the sepiapterin Form D isisolated by filtration. Typically, the sepiapterin Form D is furtherfreed from solvent (e.g., water) by drying at room temperature.

In an embodiment, the invention provides a method for preparingcrystalline Form F of sepiapterin. The method comprises preparing aslurry of sepiapterin in a solvent, wherein the solvent is water,acetone/water, isopropanol/isopropyl acetate, ortetrahydrofuran/n-hexane, and isolating sepiapterin Form D from theslurry. Preferably, the solvent is water. The sepiapterin Form D can beisolated using any suitable isolation method, for example, bycentrifugation or by filtration. Preferably, the sepiapterin Form D isisolated by filtration. Sepiapterin Form D is then converted to Form Ftypically, by heating to 40-60° C. for 0.5-10 hours. Heating may beeither at atmospheric pressure or under vacuum. Preferably heating isunder vacuum.

Sepiapterin may serve as a useful therapeutic for diseases associatedwith low intracellular BH4 levels or with dysfunction of various BH4dependent metabolic pathways including, but not limited to, primarytetrahydrobiopterin deficiency, GTPCH deficiency,6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, DHPR deficiency,sepiapterin reductase deficiency, dopamine responsive dystonia, SegawaSyndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12deficiency, Parkinson's Disease, depression due to Parkinson's Disease,impulsivity in Parkinson's patients, major depression, Autism spectrum,ADHD, schizophrenia, Bipolar disorder, cerebral ischemia, restless legsyndrome, Obsessive Compulsive Disorder, anxiety, aggression inAlzheimer's disease, cerebrovascular disorders, spasm aftersubarachnoidal hemorrhage, myocarditis, coronary vasospasm, cardiachypertrophy, arteriosclerosis, hypertension, thrombosis, infections,endotoxin shock, hepatic cirrhosis, hypertrophic pyloric stenosis,gastric mucosal injury, pulmonary hypertension, renal dysfunction,impotence, and hypoglycemia. Thus, the various forms of sepiapterin inaccordance with the present invention can be administered to a patientin an effective amount to obtain a treatment or amelioration of thedisease, disorder or condition.

The present invention further provides a pharmaceutical compositioncomprising a crystalline sepiapterin free base or a crystallinepolymorph form of a salt of sepiapterin as described above and apharmaceutically acceptable carrier. The present invention provides apharmaceutical composition comprising a pharmaceutically acceptablecarrier and an effective amount, e.g., a therapeutically effectiveamount, including a prophylactically effective amount, of one or more ofthe aforesaid compounds, or salts thereof, of the present invention.

The pharmaceutically acceptable carrier can be any of thoseconventionally used and is limited only by chemico-physicalconsiderations, such as solubility and lack of reactivity with thecompound, and by the route of administration. It will be appreciated byone of skill in the art that, in addition to the following describedpharmaceutical compositions; the compounds of the present invention canbe formulated as inclusion complexes, such as cyclodextrin inclusioncomplexes, or liposomes.

The pharmaceutically acceptable carriers described herein, for example,vehicles, adjuvants, excipients, or diluents, are well known to thosewho are skilled in the art and are readily available to the public. Itis preferred that the pharmaceutically acceptable carrier be one whichis chemically inert to the active compounds and one which has nodetrimental side effects or toxicity under the conditions of use.

The choice of carrier will be determined in part by the particularactive agent, as well as by the particular method used to administer thecomposition. Accordingly, there is a wide variety of suitableformulations of the pharmaceutical composition of the present invention.The following formulations for oral, aerosol, parenteral, subcutaneous,intravenous, intra-arterial, intramuscular, intraperitoneal,intrathecal, rectal, and vaginal administration are merely exemplary andare in no way limiting.

The crystalline sepiapterin free base or a crystalline polymorph form ofa salt of sepiapterin can be used in the preparation of liquidformulations, such as in the form of a solution, suspension, oremulsion. Formulations suitable for oral administration can consist of(a) capsules, sachets, tablets, lozenges, and troches, each containing apredetermined amount of the active ingredient, as solids or granules;(b) powders; (c) liquid solutions, such as an effective amount of thecompound dissolved in diluents, such as water, saline, or orange juice;(d) suspensions in an appropriate liquid; and (e) suitable emulsions.Preferred are solid oral dosage forms such as capsule forms, tabletforms, and powder forms. Capsule forms can be of the ordinary hard- orsoft-shelled gelatin type containing, for example, surfactants,lubricants, and inert fillers, such as lactose, sucrose, calciumphosphate, and cornstarch. Tablet forms can include one or more oflactose, sucrose, mannitol, corn starch, potato starch, alginic acid,microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicondioxide, croscarmellose sodium, talc, magnesium stearate, calciumstearate, zinc stearate, stearic acid, and other excipients, colorants,diluents, buffering agents, disintegrating agents, moistening agents,preservatives, flavoring agents, and pharmacologically compatiblecarriers. Lozenge forms can comprise the active ingredient in a flavor,usually sucrose and acacia or tragacanth, as well as pastillescomprising the active ingredient in an inert base, such as gelatin andglycerin, or sucrose and acacia, emulsions, gels, and the likecontaining, in addition to the active ingredient, such carriers as areknown in the art.

Formulations suitable for oral and/or parenteral administration includeaqueous and non-aqueous, isotonic sterile injection solutions, which cancontain anti-oxidants, buffers, bacteriostats, and solutes that renderthe formulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.The compound can be administered in a physiologically acceptable diluentin a pharmaceutical carrier, such as a sterile liquid or mixture ofliquids, including water, saline, aqueous dextrose and related sugarsolutions, an alcohol, such as ethanol, benzyl alcohol, or hexadecylalcohol, glycols, such as propylene glycol or polyethylene glycol andother polyethylene alcohols, glycerol ketals, such as2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, such as poly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester or glyceride, oran acetylated fatty acid glyceride with or without the addition of apharmaceutically acceptable surfactant, such as a soap or a detergent,suspending agent, such as pectin, carbomers, methylcellulose,hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifyingagents and other pharmaceutical adjuvants.

Oils, which can be used in parenteral formulations include petroleum,animal, vegetable, or synthetic oils. Specific examples of oils includepeanut, soybean, sesame, cottonseed, corn, olive, petrolatum, andmineral. Suitable fatty acids for use in parenteral formulations includeoleic acid, stearic acid, and isostearic acid. Ethyl oleate andisopropyl myristate are examples of suitable fatty acid esters. Suitablesoaps for use in parenteral formulations include fatty alkali metal,ammonium, and triethanolamine salts, and suitable detergents include (a)cationic detergents such as, for example, dimethyl dialkyl ammoniumhalides, and alkyl pyridinium halides, (b) anionic detergents such as,for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether,and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergentssuch as, for example, fatty amine oxides, fatty acid alkanolamides, andpolyoxyethylene-polypropylene copolymers, (d) amphoteric detergents suchas, for example, alkyl-beta-aminopropionates, and 2-alkyl-imidazopeakquaternary ammonium salts, and (3) mixtures thereof.

The parenteral formulations will typically contain from about 0.5 toabout 25% by weight of the crystalline sepiapterin free base or acrystalline polymorph form of a salt of sepiapterin in solution.Suitable preservatives and buffers can be used in such formulations. Inorder to minimize or eliminate irritation at the site of injection, suchcompositions may contain one or more nonionic surfactants having ahydrophilic-lipophilic balance (HLB) of from about 12 to about 17. Thequantity of surfactant in such formulations ranges from about 5 to about15% by weight. Suitable surfactants include polyethylene sorbitan fattyacid esters, such as sorbitan monooleate and the high molecular weightadducts of ethylene oxide with a hydrophobic base, formed by thecondensation of propylene oxide with propylene glycol. The parenteralformulations can be presented in unit-dose or multi-dose sealedcontainers, such as ampoules and vials, and can be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example, water, for injections, immediatelyprior to use. Extemporaneous injection solutions and suspensions can beprepared from sterile powders, granules, and tablets of the kindpreviously described.

The crystalline sepiapterin free base or a crystalline polymorph form ofa salt of sepiapterin of the present invention may be made intoinjectable formulations. The requirements for effective pharmaceuticalcarriers for injectable compositions are well known to those of ordinaryskill in the art. See Pharmaceutics and Pharmacy Practice, J. B.Lippincott Co., Philadelphia, Pa., Banker and Chalmers, eds., pages238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed.,pages 622-630 (1986).

Topical formulations, including those that are useful for transdermaldrug release, are well-known to those of skill in the art and aresuitable in the context of the invention for application to skin.Topically applied compositions are generally in the form of liquids,creams, pastes, lotions and gels. Topical administration includesapplication to the oral mucosa, which includes the oral cavity, oralepithelium, palate, gingival, and the nasal mucosa. In some embodiments,the composition contains at least one crystalline sepiapterin free baseor a crystalline polymorph form of a salt of sepiapterin and a suitablevehicle or carrier. It may also contain other components, such as ananti-irritant. The carrier can be a liquid, solid or semi-solid. Inembodiments, the composition is an aqueous solution. Alternatively, thecomposition can be a dispersion, emulsion, gel, lotion or cream vehiclefor the various components. In one embodiment, the primary vehicle iswater or a biocompatible solvent that is substantially neutral or thathas been rendered substantially neutral. The liquid vehicle can includeother materials, such as buffers, alcohols, glycerin, and mineral oilswith various emulsifiers or dispersing agents as known in the art toobtain the desired pH, consistency and viscosity. It is possible thatthe compositions can be produced as solids, such as powders or granules.The solids can be applied directly or dissolved in water or abiocompatible solvent prior to use to form a solution that issubstantially neutral or that has been rendered substantially neutraland that can then be applied to the target site. In embodiments of theinvention, the vehicle for topical application to the skin can includewater, buffered solutions, various alcohols, glycols such as glycerin,lipid materials such as fatty acids, mineral oils, phosphoglycerides,collagen, gelatin and silicone based materials.

The compounds of the present invention, alone or in combination withother suitable components, can be made into aerosol formulations to beadministered via inhalation. These aerosol formulations can be placedinto pressurized acceptable propellants, such asdichlorodifluoromethane, propane, nitrogen, and the like. They also maybe formulated as pharmaceuticals for non-pressured preparations, such asin a nebulizer or an atomizer.

Additionally, the crystalline sepiapterin free base or a crystallinepolymorph form of a salt of sepiapterin of the present invention may bemade into suppositories by mixing with a variety of bases, such asemulsifying bases or water-soluble bases. Formulations suitable forvaginal administration may be presented as pessaries, tampons, creams,gels, pastes, foams, or spray formulas containing, in addition to theactive ingredient, such carriers as are known in the art to beappropriate.

The crystalline sepiapterin free base or a crystalline polymorph form ofa salt of sepiapterin can be used in any suitable dose. Suitable dosesand dosage regimens can be determined by conventional range findingtechniques. Generally treatment is initiated with smaller dosages, whichare less than the optimum dose. Thereafter, the dosage is increased bysmall increments until optimum effect under the circumstances isreached. For convenience, the total daily dosage may be divided andadministered in portions during the day if desired. In proper doses andwith suitable administration of certain compounds, the present inventionprovides for a wide range of responses. Typically, the dosages rangefrom about 0.001 to about 1000 mg/kg body weight of the patient beingtreated/day. For example, in embodiments, the crystalline sepiapterinfree base or a crystalline polymorph form of a salt of sepiapterin maybe administered from about 100 mg/kg to about 300 mg/kg, from about 120mg/kg to about 280 mg/kg, from about 140 mg/kg to about 260 mg/kg, fromabout 150 mg/kg to about 250 mg/kg, from about 160 mg/kg to about 240mg/kg, of subject body weight per day, one or more times a day, toobtain the desired therapeutic effect.

In some embodiments, the crystalline sepiapterin free base or acrystalline polymorph form of a salt of sepiapterin can be formulatedinto unit solid oral dosage forms such as capsules or tablets. In theseembodiments, each unit solid oral dosage form can comprise any suitableamount of the crystalline sepiapterin free base or a crystallinepolymorph form of a salt of sepiapterin. For example, each solid oraldosage form can comprise about 10 mg, about 20 mg, about 30 mg, about 40mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg,about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg,about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg,about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg,about 475 mg, about 50 mg, about 525 mg, about 550 mg, about 575 mg,about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700 mg,about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825 mg,about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950 mg,about 975 mg, about 1000 mg, about 2000 mg, about 3000 mg, about 4000mg, about 5000 mg, and the like.

The following examples further illustrate the invention but, of course,should not be construed as in any way limiting its scope.

Abbreviations: MeOH—methanol; DMSO—dimethylsulfoxide; EtOAc—ethylacetate; DMAc—dimethylacetamide; THF—tetrahydrofuran;NMP—N-methylpyrrolidone.

For X-ray powder diffraction analysis, a PANalytical Empyrean X-raypowder diffractometer was used. The parameters are as follows:

XRD Parameters

Parameter Value X-Ray wavelength Cu, kα, Kα1 (Å): 1.540598, Kα2 (Å):1.544426 Kα2/Kα1 intensity ratio: 0.50 X-Ray tube setting 45 kV, 40 mADivergence slit Automatic Scan mode Continuous Scan range (°2θ) 3°~40°Scan step time (s) 17.8 Test time (s) 5 min 30 s

DSC was performed using a TA Q200/Q2000 DSC from TA Instruments.Parameters used are as follows:

Parameters DSC Method Ramp Sample pan Aluminum, crimped Temperature 25°C. - desired temperature Heating rate 10° C./min Purge gas N₂

EXAMPLES

While certain features of the invention have been illustrated anddescribed herein, many modifications, substitutions, changes, andequivalents will now occur to those of ordinary skill in the art. It is,therefore, to be understood that the appended claims are intended tocover all such modifications and changes as fall within the true spiritof the invention. As such, the following examples are provided to teachvarious aspects of the present invention. These examples representindividual embodiments of the aspects of this invention and one skilledin the art will recognize that additional examples can be generated inorder to equally teach the aspects of the present invention.

Example 1

This example demonstrates a preparation of the crystalline Form B ofsepiapterin free base in accordance with an embodiment of the invention.

73.2 mg of starting material sepiapterin was weighed into a 20-mL glassvial. 2.5 mL of N-methyl pyrrolidone (NMP) was added to dissolve thestarting material. The solution was filtered into a new vial. 17 mL ofacetonitrile (ACN) was added step-wise, with the sample stirring at RTwith a rate of ˜1000 rpm. The suspension was stirred at RT for 2 hrs.The resulting precipitate was isolated by centrifugation and dried invacuum at RT for 3 hrs to obtain crystalline Form B of sepiapterin freebase.

Example 2

This example demonstrates a preparation of crystalline Form C ofsepiapterin free base in accordance with an embodiment of the invention.

100.4 mg of starting material sepiapterin was weighed into a 20-mL glassvial. 2 mL of ACN was added to form a suspension, which was stirred at50° C. with a rate of ˜1000 rpm. The resulting solids were isolated bycentrifugation for 2 minutes through a 0.25 □m pore size centrifugationfilter and drying at RT for approximately 12 hours to obtain crystallineForm C of sepiapterin free base.

Example 3

This example demonstrates a preparation of the crystalline Form D ofsepiapterin free base in accordance with an embodiment of the invention.

200.1 mg of starting material sepiapterin was weighed into a 20-mL glassvial. 5 mL of H₂O was added to form a suspension, which was stirred at50° C. with a rate of ˜1000 rpm. The resulting solids were isolated bycentrifugation for 2 minutes through a 0.25 □m pore size centrifugationfilter. One-half of the collected solids were dried at RT forapproximately 12 hours at atmospheric pressure to obtain crystallineForm D of sepiapterin free base.

Example 4

This example demonstrates a preparation of the crystalline Form F ofsepiapterin free base in accordance with an embodiment of the invention.The other half of the collected solids from Example 3 were dried undervacuum at 50° C. for 0.5 hr to obtain crystalline Form F of sepiapterinfree base.

Example 5

This example demonstrates a preparation of the crystalline Form G ofsepiapterin free base in accordance with an embodiment of the invention.The crystalline Form G of sepiapterin free base was prepared by heatinga sample of crystalline Form F prepared as in Example 4 to 120° C. underN₂ flow.

Example 6

This example demonstrates a preparation of the crystalline hydrochloridesalt of sepiapterin free base in accordance with an embodiment of theinvention.

120.4 mg of sepiapterin freebase was weighed into a 20-mL glass vial.0.8 mL of acetone/H₂O (9:1, v/v) and 42 μL of conc. HCl (37.5%) wereadded, and the resulting suspension was stirred at RT at a rate of ˜1000rpm for 5 days. The resulting solids were isolated by vacuum filtrationand dried in vacuum at RT for 3 hrs.

The solids obtained above were dispersed in 3 mL of acetone/H₂O (9:1,v/v). 5.5 μL of conc. HCl (37.5%) was added and the suspension wasstirred at RT at a rate of ˜1000 rpm for 6 days, following which, thesolids were isolated by vacuum filtration and dried in vacuum at RTovernight to obtain the crystalline hydrochloride salt of sepiapterinfree base.

Example 7

This example demonstrates a preparation of the crystalline Form 3methanesulfonate salt of sepiapterin free base in accordance with anembodiment of the invention.

51.7 mg of methanesulfonic acid was weighed into a 20-mL glass vial. 5mL of MeOH was added to the vial. 120.7 mg of sepiapterin freebase wasweighed into the vial. The resulting suspension was stirred at RT at arate of ˜1000 rpm for 5 days, following which 20 μL of methanesulfonicacid was added to the vial. The resulting mixture was stirred at RT at arate of ˜1000 rpm for 1 day. The solids were isolated by vacuumfiltration and dried in vacuum at RT overnight. The dried solids weredispersed in 3 mL of MeOH and stirred at RT at a rate of ˜1000 rpm for 1day. The solids were isolated by vacuum filtration and dried in vacuumat RT overnight to obtain crystalline Form 3 methanesulfonate salt ofsepiapterin free base.

Example 8

This example demonstrates a preparation of the crystalline nicotinatesalt of sepiapterin free base in accordance with an embodiment of theinvention.

119.5 mg of freebase was weighed into a 20-mL glass vial. 10 mL of MeOHwas added to the vial. 100.1 mg of nicotinic acid was weighed into thevial. The resulting suspension was stirred at RT at a rate of ˜1000 rpmfor 7 hrs, following which the obtained solids were isolated by vacuumfiltration and dried in vacuum at RT for 3 hrs to obtain the crystallinenicotinate salt of sepiapterin free base.

Example 9

This example demonstrates a preparation of the crystalline salt forms ofsepiapterin free base in accordance with an embodiment of the invention.

Crystalline form 1 sulfate salt was obtained by slurrying equimolaramounts of starting material and H₂SO₄ in acetone/H₂O (9:1, v/v).

Crystalline form 2 sulfate salt was obtained by slurrying equimolaramounts of starting material and H₂SO₄ in THF/DMAc (9:1, v/v).

Crystalline p-toluenesulfonate salt was obtained by slurrying equimolaramounts of starting material and p-toluene sulfonic acid in methanol.

Crystalline Form 1 methanesulfonate salt was obtained by slurryingequimolar amounts of starting material and methane sulfonic acid inmethanol.

Crystalline Form 2 methanesulfonate salt was obtained by slurryingequimolar amounts of starting material and methane sulfonic acid inacetone/H₂O (9:1, v/v).

Crystalline benzenesulfonate salt was obtained by slurrying equimolaramounts of starting material and benzene sulfonic acid in methanol.

Crystalline phosphate salt was obtained by slurrying equimolar amountsof starting material and H₃PO₄ in acetone/H₂O (9:1, v/v).

Crystalline malonate salt was obtained by slurrying starting materialand malonic acid (molar ratio of acid/freebase about 5:1) in acetone/H₂O(9:1, v/v).

Crystalline L-tartrate salt was obtained by slurrying starting materialand gentisic acid (molar ratio of acid/freebase about 4:1) inacetone/H₂O (9:1, v/v).

Crystalline gentisate salt was obtained by slurrying starting materialand L-tartaric acid (molar ratio of acid/freebase about 5:1) inacetone/H₂O (9:1, v/v).

Crystalline fumarate salt was obtained by slurrying starting materialand fumaric acid (molar ratio of acid/freebase about 5:1) in acetone/H₂O(9:1, v/v).

Crystalline glycolate salt was obtained by slurrying starting materialand glycolic acid (molar ratio of acid/freebase about 4:1) inacetone/H₂O (9:1, v/v).

Crystalline acetate salt was obtained by slurrying starting material andacetic acid (molar ratio of acid/freebase about 5:1) in acetone/H₂O(9:1, v/v).

Example 10

This example demonstrates characterization of the starting sepiapterinused in the preparation of the crystalline polymorphs A, B, C, D, E, F,and G of sepiapterin free base and of the crystalline polymorph forms ofsalts of sepiapterin described herein.

A sample of sepiapterin free base was obtained commercially. DSC showedtwo endotherms at 82.8° C. and 179.8° C. The sepiapterin samplecontained particles with an average particle size over 100 μm. The XRDpattern was determined before and after grinding to reduce the particlesize such that it passes through a 140 mesh screen. The XRD patternsboth before and after grinding are shown in FIG. 19. This polymorph ofsepiapterin free base is referred to as Form A herein.

Example 11

This example demonstrates the results of stability studies carried outon the sepiapterin starting material (Form A), crystalline polymorphForm D, and crystalline polymorph Form F at temperatures of roomtemperature (RT), 35° C., and 50° C.

The purity of initial samples was determined by HPLC and was found to beas follows: Form A=99.3 area %, Form F=99.7 area %, Form D=99.1 area %,wherein area % refers to the area under the curve of the sepiapterinpeak as compared with the total area under all of the peaks.

Form A and F samples were placed in chambers with silica gel to removewater (the relative humidity was measured to be ˜10% RH) at differenttemperatures. Form D samples were placed in chambers with water(relative humidity was estimated to be ˜100% RH) at differenttemperatures.

The HPLC purities and XRD patterns were obtained for each of Form A/F/Dsamples stored at various temperatures. The results after 1 week and 4weeks of storage are set forth in Tables 22 and 23, respectively.

TABLE 24 Storage after 1 week RT 35° C 50° C Purity % of Form Purity %of Form Purity % of Form Sample Humidity (area %) Initial change (area%) Initial change (area %) Initial change Form A 10% RH 98.8 99.5 No98.0 98.6 No 95.2 95.8 No Form F 10% RH 99.4 99.7 No 99.4 99.7 No 99.299.4 No Form D 100% RH 99.0 99.9 No 98.8 99.7 No 98.5 99.3 No

TABLE 25 Storage after 4 weeks RT 35° C 50° C Purity % of Form Purity %of Form Purity % of Form Sample Humidity (area %) Initial change (area%) Initial change (area %) Initial change Form A 10% RH 98.2 98.8 No96.1 96.7 No 88.5 89.1 No Form F 10% RH 99.5 99.8 No 99.4 99.7 No 98.999.1 No Form D 100% RH 98.9 99.8 No 98.7 99.6 No 97.7 98.5 No

As is apparent from the results set forth in Tables 24 and 25, none ofthe samples exhibited a significant change in crystal structure asobserved by XPD. Form A exhibited significantly less stability asdetermined by HPLC. After storage for 4 weeks at 50° C., the purity ofForm A as measured by HPLC peak area % was 89.1% compared to the initialpurity. The purities of Forms F and D were 99.1% and 98.5%,respectively, compared to the initial purity.

Example 12

This example demonstrates the stability of polymorphs D and F ofsepiapterin free base on storage.

Samples of sepiapterin free base polymorph Forms D, F, and A were storedat room temperature (RT), 35° C., and 50° C. The samples were analyzedby HPLC at 1 week and 4 week intervals. The HPLC parameters were asfollows:

Parameters Solubility Stability (purity) Column Inertsil ODS-3, 4.6 ×250 mm, 5 μm Mobile phase A:20 mM K₂HPO₄—KH₂PO₄ buffer (pH 7.0):ACN(98:2) B:20 mM K₂HPO₄—KH₂PO₄ buffer (pH 7.0):ACN (50:50) Time (min) % BTime (min) % B Gradient table 0.0 0 0.0 0 3.0 0 5.0 0 10.0 100 25.0 10010.1 0 25.1 0 12.0 0 35.0 0 Run time 12.0 min 35.0 min Post time 0.0 minFlow rate 1.0 mL/min Injection volume 5 μL Detector wavelength UV at 280nm Column temperature 40° C. Sampler temperature RT Diluent H₂O

The results for polymorphs A, F, and D of sepiapterin free base are setforth in Tables 26-28.

TABLE 26 Polymorph A RT/10% RH 35° C./10% RH 50° C./10% RH # RRT Initial1 w 4 w 1 w 4 w 1 w 4 w Impurity 0.62 0.08 0.16 0.37 0.24 1.03 0.68 2.59Impurity 0.89 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 0.06 Impurity 0.950.42 0.77 1.29 1.36 2.70 3.48 7.67 Sepiapterin 1.00 99.33 98.80 98.1897.97 96.06 95.15 88.49 Impurity 1.06 <0.05 <0.05 <0.05 <0.05 <0.05 0.45<0.05 Impurity 1.08 <0.05 0.11 <0.05 0.28 <0.05 0.08 0.85 Impurity 1.170.17 0.16 0.16 0.16 0.16 0.16 0.17 Impurity 1.21 <0.05 <0.05 <0.05 <0.05<0.05 <0.05 0.06 Impurity 1.25 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 0.11

TABLE 27 Polymorph F RT/10% RH 35° C./10% RH 50° C./10% RH # RRT Initial1 w 4 w 1 w 4 w 1 w 4 w Impurity 0.60 <0.05 <0.05 <0.05 <0.05 <0.05<0.05 0.12 Impurity 0.95 0.09 0.29 0.30 0.35 0.42 0.58 0.86 Sepiapterin1.00 99.74 99.42 99.54 99.43 99.43 99.15 98.85 Impurity 1.08 <0.05 0.14<0.05 0.06 <0.05 0.11 <0.05 Impurity 1.17 0.17 0.16 0.16 0.15 0.15 0.150.17

TABLE 28 Polymorph D RT/100% RH 35° C./100% RH 50° C./100% RH # RRT*Initial 1 w 4 w 1 w 4 w 1 w 4 w Impurity 0.62 <0.05 <0.05 0.09 <0.050.09 0.08 0.14 Impurity 0.95 0.69 0.83 0.81 0.86 1.04 1.05 2.03Sepiapterin 1.00 99.14 99.01 98.92 98.83 98.70 98.45 97.65 Impurity 1.08<0.05 <0.05 <0.05 0.14 <0.05 0.26 <0.05 Impurity 1.17 0.17 0.16 0.170.17 0.17 0.16 0.18 *—Relative retention time

As is apparent from the results set forth in Tables 26-28, polymorphs Dand F of sepiapterin free base exhibited significantly greater stabilitythan polymorph A. The amount of sepiapterin in polymorph A decreasedfrom 99.33% for 88.49% after storage for 4 weeks at 50° C./10% RH(relative humidity). The amount of sepiapterin in polymorph D decreasedfrom 99.14% to 97.65% after storage for 4 weeks at 50° C./100% RH. Theamount of sepiapterin in polymorph F decreased from 99.74% to 98.85%after storage for 4 weeks at 50° C./10% RH.

Example 13

This example demonstrates a preparation of crystalline Form E ofsepiapterin free base.

100.6 mg of starting material was weighed into a 3-mL glass vial. 1 mLof MeOH was added to form a suspension. The sample was stirred at RTwith a rate of ˜1000 rpm. The resulting solids were isolated bycentrifugation after 3 days and dried at RT overnight.

Other Embodiments

The use of the terms “a” and “an” and “the” and “at least one” andsimilar referents in the context of describing the invention (especiallyin the context of the following claims) are to be construed to coverboth the singular and the plural, unless otherwise indicated herein orclearly contradicted by context. The use of the term “at least one”followed by a list of one or more items (for example, “at least one of Aand B”) is to be construed to mean one item selected from the listeditems (A or B) or any combination of two or more of the listed items (Aand B), unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof any and all examples, or exemplary language (e.g., “such as”)provided herein, is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention unlessotherwise claimed. No language in the specification should be construedas indicating any non-claimed element as essential to the practice ofthe invention.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Variations of those preferred embodiments may become apparent to thoseof ordinary skill in the art upon reading the foregoing description. Theinventors expect skilled artisans to employ such variations asappropriate, and the inventors intend for the invention to be practicedotherwise than as specifically described herein. Accordingly, thisinvention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

What is claimed:
 1. A crystalline form sepiapterin having at least onepeak at diffraction angle 2θ (°) of 8.4°±0.5, 16.9°±0.5, or 25.4°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry.
 2. The crystalline form ofsepiapterin of claim 1 having at least one peak at diffraction angle 2θ(°) of 8.4°±0.5, 16.9°±0.5, and 25.4°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 3. The crystalline form of sepiapterin of claim 1 or 2having at least one peak at diffraction angle 2θ (°) of 14.9°±0.5, or34.1°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry.
 4. The crystalline formof sepiapterin of any one of claims 1 to 3 having at least one peak atdiffraction angle 2θ (°) of 8.4°±0.5, 14.9°±0.5, 16.9°±0.5, 25.4°±0.5,and 34.1°±0.5 as measured by X-ray diffractometry by irradiation with CuKα X-rays or calculated from X-ray diffractometry.
 5. The crystallineform of sepiapterin of any one of claims 1 to 4 having the X-ray powderdiffraction spectrum as shown in FIG.
 1. 6. The crystalline form ofsepiapterin of any one of claims 1 to 5 having an endothermic onset atabout 195° C. in differential scanning calorimetry (DSC) profile.
 7. Acrystalline form sepiapterin having at least one peak at diffractionangle 2θ (°) of 5.7°±0.5, 7.8°±0.5, or 25.4°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 8. The crystalline form of sepiapterin of claim 7 havingat least one peak at diffraction angle 2θ (°) of 5.7°±0.5, 7.8°±0.5, and25.4°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry.
 9. The crystalline formof sepiapterin of claim 7 or 8 having at least one peak at diffractionangle 2θ (°) of 9.1°±0.5, 11.5°±0.5, 15.3°±0.5, 16.0°±0.5, 20.1°±0.5, or26.6°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry.
 10. The crystalline formof sepiapterin of any one of claims 7 to 9 having at least one peak atdiffraction angle 2θ (°) of 5.7°±0.5, 7.8°±0.5, 9.1°±0.5, 11.5°±0.5,15.3°±0.5, 16.0°±0.5, 20.1°±0.5, 25.4°±0.5, and 26.6°±0.5 as measured byX-ray diffractometry by irradiation with Cu Kα X-rays or calculated fromX-ray diffractometry.
 11. The crystalline form of sepiapterin of any oneof claims 7 to 10 having the X-ray powder diffraction spectrum as shownin FIG.
 2. 12. The crystalline form of sepiapterin of any one of claims7 to 11 having an endothermic onset at about 58° C., 102° C., 130° C.,156.5° C., or 168° C. in differential scanning calorimetry (DSC)profile.
 13. The crystalline form of sepiapterin of any one of claims 7to 12 having an endothermic onset at about 58° C., 102° C., 130° C.,156.5° C., and 168° C. in differential scanning calorimetry (DSC)profile.
 14. A crystalline form sepiapterin having at least one peak atdiffraction angle 2θ (°) of 8.9°±0.5, 10.3°±0.5, or 26.0°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry.
 15. The crystalline form ofsepiapterin of claim 14 having at least one peak at diffraction angle 2θ(°) of 8.9°±0.5, 10.3°±0.5, and 26.0°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 16. The crystalline form of sepiapterin of claim 14 or15 having at least one peak at diffraction angle 2θ (°) of 10.9°±0.5,17.8°±0.5, 24.9°±0.5, 26.7°±0.5, 26.8°±0.5, or 28.3°±0.5 as measured byX-ray diffractometry by irradiation with Cu Kα X-rays or calculated fromX-ray diffractometry.
 17. The crystalline form of sepiapterin of any oneof claims 14 to 16 having at least one peak at diffraction angle 2θ (°)of 8.9°±0.5, 10.3°±0.5, 10.9°±0.5, 17.8°±0.5, 24.9°±0.5, 26.0°±0.5,26.7°±0.5, 26.8°±0.5, and 28.3°±0.5 as measured by X-ray diffractometryby irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 18. The crystalline form of sepiapterin of any one ofclaims 14 to 17 having the X-ray powder diffraction spectrum as shown inFIG.
 3. 19. The crystalline form of sepiapterin of any one of claims 14to 18 having an endothermic onset at about 43° C., 66° C., or 233° C. indifferential scanning calorimetry (DSC) profile.
 20. The crystallineform of sepiapterin of any one of claims 14 to 19 having an endothermiconset at about 43° C., 66° C., and 233° C. in differential scanningcalorimetry (DSC) profile.
 21. A crystalline form sepiapterin having atleast one peak at diffraction angle 2θ (°) of 9.7°±0.5, 10.2°±0.5, or11.3°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry.
 22. The crystalline formof sepiapterin of claim 21 having at least one peak at diffraction angle2θ (°) of 9.7°±0.5, 10.2°±0.5, and 11.3°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 23. The crystalline form of sepiapterin of claim 21 or22 having at least one peak at diffraction angle 2θ (°) of 14.0°±0.5,14.6°±0.5, 19.9°±0.5, 22.2°±0.5, 25.3°±0.5, or 32.4°±0.5 as measured byX-ray diffractometry by irradiation with Cu Kα X-rays or calculated fromX-ray diffractometry.
 24. The crystalline form of sepiapterin of any oneof claims 21 to 23 having at least one peak at diffraction angle 2θ (°)of 9.7°±0.5, 10.2°±0.5, 11.3°±0.5, 14.0°±0.5, 14.6°±0.5, 19.9°±0.5,22.2°±0.5, 25.3°±0.5, and 32.4°±0.5 as measured by X-ray diffractometryby irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 25. The crystalline form of sepiapterin of any one ofclaims 21 to 24 having the X-ray powder diffraction spectrum as shown inFIG.
 4. 26. The crystalline form of sepiapterin of any one of claims 21to 25 having an endothermic onset at about 113° C. or 196° C. indifferential scanning calorimetry (DSC) profile.
 27. The crystallineform of sepiapterin of any one of claims 21 to 26 having an endothermiconset at about 113° C. and 196° C. in differential scanning calorimetry(DSC) profile.
 28. A crystalline form sepiapterin having at least onepeak at diffraction angle 2θ (°) of 10.0°±0.5, 10.6°±0.5, or 25.7°±0.5as measured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry.
 29. The crystalline form ofsepiapterin of claim 28 having at least one peak at diffraction angle 2θ(°) of 10.0°±0.5, 10.6°±0.5, and 25.7°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 30. The crystalline form of sepiapterin of claim 28 or29 having at least one peak at diffraction angle 2θ (°) of 11.2°±0.5,15.3°±0.5, 15.9°±0.5, 22.8°±0.5, 24.4°±0.5, or 25.0°±0.5 as measured byX-ray diffractometry by irradiation with Cu Kα X-rays or calculated fromX-ray diffractometry.
 31. The crystalline form of sepiapterin of any oneof claims 28 to 30 having at least one peak at diffraction angle 2θ (°)of 10.0°±0.5, 10.6°±0.5, 11.2°±0.5, 15.3°±0.5, 15.9°±0.5, 22.8°±0.5,24.4°±0.5, 25.0°±0.5, and 25.7°±0.5 as measured by X-ray diffractometryby irradiation with Cu Kα X-rays or calculated from X-raydiffractometry.
 32. The crystalline form of sepiapterin of any one ofclaims 28 to 31 having the X-ray powder diffraction spectrum as shown inFIG.
 5. 33. A composition comprising a crystalline form of sepiapterinof any one of claims 1 to
 32. 34. The composition of claim 33, wherein acrystalline form of sepiapterin of any one of claims 1 to 27, is presentin an amount of at least 90 percent by weight of the composition.
 35. Apharmaceutical composition comprising the crystalline form ofsepiapterin of any one of claims 1 to 32 and a pharmaceuticallyacceptable carrier.
 36. The pharmaceutical composition of claim 35,wherein the crystalline form of sepiapterin is formulated as particlesless than 100 μm in size.
 37. A method for preparing a crystalline formof sepiapterin comprising preparing a slurry of a first crystalline formof sepiapterin in water, acetone/water, isopropanol/isopropyl acetate,or tetrahydrofuran/n-hexane, isolating the solids from the slurry, anddrying the solids.
 38. The method of claim 37, wherein the slurry of thefirst crystalline form of sepiapterin is stirred at 25-75° C. for 6-72hours.
 39. The method of claim 37 or 38, wherein the solids are dried at20-30° C. for 6-24 hours.
 40. The method of claim 37 or 38, wherein thesolids are dried at 40-60° C. for 5-10 hours.
 41. The method of any oneof claims 37 to 40, wherein the solids are dried at atmosphericpressure.
 42. The method of any one of claims 37 to 40, wherein thesolids are dried under vacuum.
 43. A crystalline form of a salt ofsepiapterin, wherein the crystalline form of a salt of sepiapterin is:(a) a crystalline form of the methanesulfonate salt of sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 7.8°±0.5,23.5°±0.5, and 29.0°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry;(b) a crystalline form of the methanesulfonate salt of sepiapterinhaving at least one peak at diffraction angle 2θ (°) of 21.7°±0.5,26.0°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry(c) a crystalline form of the nicotinate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 9.5°±0.5, 9.9°±0.5, and24.5°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry; (d) a crystalline formof the p-toluenesulfonate salt of sepiapterin having at least one peakat diffraction angle 2θ (°) of 6.5°±0.5, 15.1°±0.5, and 23.4°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry; (e) a crystalline form of thebenzenesulfonate salt of sepiapterin having at least one peak atdiffraction angle 2θ (°) of 6.5°±0.5, 14.8°±0.5, and 19.6°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry; (f) a crystalline form of thephosphate salt of sepiapterin having at least one peak at diffractionangle 2θ (°) of 16.6°±0.5, 22.2°±0.5, and 25.6°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry; (g) a crystalline form of the malonate salt ofsepiapterin having at least one peak at diffraction angle 2θ (°) of6.9°±0.5, 22.7°±0.5, and 23.8°±0.5 as measured by X-ray diffractometryby irradiation with Cu Kα X-rays or calculated from X-raydiffractometry; (h) a crystalline form of the tartrate salt ofsepiapterin having at least one peak at diffraction angle 2θ (°) of7.3°±0.5, 14.2°±0.5, and 21.8°±0.5 as measured by X-ray diffractometryby irradiation with Cu Kα X-rays or calculated from X-raydiffractometry; (i) a crystalline form of the gentisate salt ofsepiapterin having at least one peak at diffraction angle 2θ (°) of7.1°±0.5, 8.7°±0.5, and 26.7°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry;(j) a crystalline form of the fumarate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 11.3°±0.5, 24.0°±0.5, and28.2°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry; (k) a crystalline formof the glycolate salt of sepiapterin having at least one peak atdiffraction angle 2θ (°) of 7.6°±0.5, 10.7°±0.5, and 24.0°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry; (l) a crystalline form of theacetate salt of sepiapterin having at least one peak at diffractionangle 2θ (°) of 6.2°±0.5, 12.0°±0.5, and 18.1°±0.5 as measured by X-raydiffractometry by irradiation with Cu Kα X-rays or calculated from X-raydiffractometry; (m) a crystalline form of the sulfate salt ofsepiapterin having at least one peak at diffraction angle 2θ (°) of5.1°±0.5, 7.8°±0.5, and 23.0°±0.5 as measured by X-ray diffractometry byirradiation with Cu Kα X-rays or calculated from X-ray diffractometry;or (n) a crystalline form of the sulfate salt of sepiapterin having atleast one peak at diffraction angle 2θ (°) of 7.8°±0.5, 8.8°±0.5, and24.1°±0.5 as measured by X-ray diffractometry by irradiation with Cu KαX-rays or calculated from X-ray diffractometry.
 44. A crystalline formof the hydrochloride salt of sepiapterin having at least one peak atdiffraction angle 2θ (°) of 7.8°±0.5, 12.9°±0.5, and 26.2°±0.5 asmeasured by X-ray diffractometry by irradiation with Cu Kα X-rays orcalculated from X-ray diffractometry
 45. A composition comprising acrystalline form of a salt of sepiapterin of claim 43 or 44, wherein thecrystalline form of the salt of sepiapterin is present in at least 90percent by weight.
 46. A pharmaceutical composition comprising thecrystalline form of a salt of sepiapterin of claim 43 or 44 and apharmaceutically acceptable carrier.
 47. The pharmaceutical compositionof claim 46, wherein the crystalline form of sepiapterin is formulatedas particles less than 100 μm in size.
 48. A method for treating a BH4related disorder in a patient in need thereof, the method comprisingadministering to the patient an effective amount of the crystalline formof sepiapterin of any one of claim 1-32, 43, or 44, or a pharmaceuticalcomposition of any one of claim 35, 36, 46, or
 47. 49. A method ofincreasing BH4 levels in a subject in need thereof, the methodcomprising administering to the patient an effective amount of thecrystalline form of sepiapterin of any one of claim 1-32, 43, or 44, ora pharmaceutical composition of any one of claim 35, 36, 46, or
 47. 50.A method of decreasing phenylalanine levels in a subject in needthereof, the method comprising administering to the patient an effectiveamount of the crystalline form of sepiapterin of any one of claim 1-32,43, or 44, or a pharmaceutical composition of any one of claim 35, 36,46, or
 47. 51. A method of increasing the activity of phenylalaninehydroxylase in a subject, the method comprising administering to thepatient an effective amount of the crystalline form of sepiapterin ofany one of claim 1-32, 43, or 44, or a pharmaceutical composition of anyone of claim 35, 36, 46, or
 47. 52. A method of treating phenylketonuriain a subject in need thereof, the method comprising administering to thepatient an effective amount of the crystalline form of sepiapterin ofany one of claim 1-32, 43, or 44, or a pharmaceutical composition of anyone of claim 35, 36, 46, or
 47. 53. A method of increasing serotoninlevels in a subject in need thereof, the method comprising administeringto the patient an effective amount of the crystalline form ofsepiapterin of any one of claim 1-27, 38, or 39, or a pharmaceuticalcomposition of any one of claim 30, 31, 41, or
 42. 54. A method ofincreasing the activity of tryptophan hydroxylase in a subject, themethod comprising administering to the patient an effective amount ofthe crystalline form of sepiapterin of any one of claim 1-27, 38, or 39,or a pharmaceutical composition of any one of claim 30, 31, 41, or 42.55. A method of increasing dopamine levels in a subject in need thereof,the method comprising administering to the patient an effective amountof the crystalline form of sepiapterin of any one of claim 1-27, 38, or39, or a pharmaceutical composition of any one of claim 30, 31, 41, or42.
 56. A method of increasing the activity of tyrosine hydroxylase in asubject, the method comprising administering to the patient an effectiveamount of the crystalline form of sepiapterin of any one of claim 1-27,38, or 39, or a pharmaceutical composition of any one of claim 30, 31,41, or
 42. 57. A method of increasing the activity of nitric oxidesynthases in a subject, the method comprising administering to thepatient an effective amount of the crystalline form of sepiapterin ofany one of claim 1-27, 38, or 39, or a pharmaceutical composition of anyone of claim 30, 31, 41, or
 42. 58. A method of increasing the activityof alkylglycerol monooxygenase in a subject, the method comprisingadministering to the patient an effective amount of the crystalline formof sepiapterin of any one of claim 1-27, 38, or 39, or a pharmaceuticalcomposition of any one of claims 30, 31, 41, or 42.